Compounds and compositions for reducing lipid levels

ABSTRACT

Compositions comprising extracts or isolated or purified compounds from plants of the genus  Corydalis  provide prevention and treatment of hyperlipidemia, hypercholesterolemia, hypertriglyceridemia, hepatic steatosis, and metabolic syndrome.  Corydalis  compounds and their derivatives of natural and synthetic origins lower total cholesterol, LDL-cholesterol, and triglycerides and increase hepatic LDL receptor expression and activate AMP-activated protein kinase. Specific stereoisomers of  Corydalis  compounds with lipid lowering activity include 14R-(+)-corypalmine, 14R,13S-(+)-corydaline, 14R-(+)-tetrahydropalmatin, (+)-corlumidin, d-(+)-bicuculline, and (+)-egenine.

RELATED APPLICATIONS

The present application claims priority to U.S. Provisional PatentApplication No. 60/937,007, filed Jun. 22, 2007, and to U.S. ProvisionalPatent Application No. 61/054,444, filed May 19, 2008, the entirecontents of which are incorporated by reference herein.

FIELD OF THE INVENTION

The present invention is related to methods, compounds and compositionsto treat hyperlipidemia, including hypertriglyceridemia andhypercholesterolemia, as well as hepatic steatosis and metabolicsyndrome.

SUMMARY OF THE INVENTION

In accordance with one aspect, the invention provides methods ofreducing plasma and/or hepatic lipid levels of a subject in needthereof, which comprises administering to the said subject alipid-lowering effective amount of a compound, composition or extractdescribed herein. The lipid level to be reduced can be one or more oftotal cholesterol, LDL-cholesterol, triglycerides, and unesterified longchain fatty acids. In another aspect, the invention provides methods fortreating a disease or condition selected from the group consisting ofhyperlipidemia, hypercholesterolemia, hypertriglyceridemia, hepaticsteatosis and metabolic syndrome, comprising administering to a subjectin need thereof a therapeutically effective amount of a compound,composition or extract described herein.

In one aspect, the invention provides lipid lowering agents, includinghypocholesterolemic and/or hypotriglyceridemic extracts and compounds,and derivatives of such compounds, from a variety of plants includingCorydalis, Leontice, Mahonia, Fumaria, Legnephora, Stephania,Chelidonium, Hunnemannia, Coptis, Guatteria, Pachypodanthium;Chasmanthera, Fibraurea; Cheilanthes, Dicranostigma; Glaucium; andChelidonium. In some embodiments, the extract is obtained from the plantspecies selected from the group consisting of Corydalis (ambigua,bulbosa, cava, chaerophylla, pallida, solida, thalictrifolia., tuberosa,turtschaminowii Besser), Leontice (leontopetalum), Mahonia (aquifolium),Fumaria (vaillantii), Legnephora (moorii), Stephania (glabra, tetranda),Chelidonium (majus), Hunnemannia (fumariaefolia), Coptis (groenlandica),Guatteria (discolor), Pachypodanthium (staudtii); Chasmanthera(dependens), Fibraurea (chloroleuca); Cheilanthes (meifolia),Dicranostigma. (leptopodum); Glaucium (vitellinum); Corydalis yan husuo; and Corydalis Xiar Ri Wu.

In certain embodiments the lipid lowering agent isisoquinolinyl-containing alkaloid from, e.g., a Corydalis extract or aderivative of a Corydalis compound, such as a compound of Formulae I,II, III, or IV as shown herein. Exemplary lipid lowering agents includesubstantially pure corlumidin (CLMD), (+)-corlumidin, (+)-CLMD,corypalmine (CRPM), 14R-(+)-corypalmine (14R-(+)-CRPM),tetrahydropalmatine (THP), 14R-(+)-tetrahydropalmatine (14R-(+)-THP),corydaline (CRDL), 14R,13S-(+)-corydaline (14R,13S-(+)-CRDL),bicuculline (BCCL), d-(+)-bicuculline (d-(+)-BCCL), and Egenine (EGN),(+)-egenine ((+)-EGN).

For compounds of either Formula I or Formula II, R₁, R₂, R₃, R₄, R₅, andR₆ are selected (independently, collectively, or in any combination)from H, halogen, hydroxy, C₁-C₆ alkyl, alkoxy, nitro, amino, aminoalkyl,trifluoromethyl, trifluoromethoxy, cycloalkyl, alkanoyl, alkanoyloxy,nitrile, dialkylamino, alkenyl, hydroxyalkyl, alkylaminoalkyl,aminoalkyl, dialkylaminoalkyl, haloalkyl, carboxyalkyl, alkoxyalkyl,carboxy, alkanoylamino, carbonylamino, carbamoyl, alkylsulfonylamino,and heterocyclyl groups. Preferably, R₁, R₂, R₃, R₄, R₅, and R₆ are nothalogen when halogen would be covalently bonded to oxygen. In oneaspect, the compounds of the invention can also comprise one or morehalogens as substituents at any position of Formula I or Formula II. Insome embodiments, compounds of Formula I have the 14R-(+) stereochemicalconfiguration.

In some embodiments, the lipid lowering agents that may be used inmethods described herein include compounds of Formula III and FormulaIV:

or stereoisomers thereof, tautomers thereof, solvates thereof, andpharmaceutically acceptable salts thereof; wherein

R₁ and R₂ are independently —H, —(CH₂)₀₋₆COOR′, —C(O)R″, or asubstituted or unsubstituted alkyl, cycloalkyl, cycloalkylalkyl,alkenyl, aryl, aralkyl, heteroaryl, heteroarylalkyl, heterocyclyl, orheterocyclylalkyl group; or R₁ and R₂ together are a methylene group;

R₃ and R₈ are independently —H, —OH, —Cl, —Br, —F, —I, —CN, —NH₂,—C(O)NH₂, —COOH, or a substituted or unsubstituted alkyl, alkoxy,alkenyl, or aralkyl group;

R₃′ is H, or R₃ and R₃′ together are an oxo group;

R₄ is —H, halogen, —OR′, —OSO₂R″, —OC(O)R″, —OC(O)OR″, —OC(O)NR′R″,—O-alkylene-NR′R′, —O-alkylene-OSO₂R″, —O-alkylene-S(O)₀₋₂R″,—O-alkylene-NR′SO₂R″, —O-alkylene-N(R′)C(O)R′, or a substituted orunsubstituted alkyl group;

R₅ and R₆ are independently —H, halogen, —OH, or a substituted orunsubstituted alkoxy group; or R₄ and R₅ together are a methylenedioxygroup, or R₅ and R₆ together are a methylenedioxy group;

R₇ is H, halogen, OH, or a substituted or unsubstituted alkyl or alkoxygroup;

R₉ is H or a substituted or unsubstituted alkyl group;

each R′ is independently a hydrogen, or a substituted or unsubstitutedalkyl, alkenyl, cycloalkyl, cycloalkylalkyl, aryl, aralkyl, heteroaryl,heteroarylalkyl, heterocyclyl, or heterocyclylalkyl group;

each R″ is independently a substituted or unsubstituted alkyl, alkenyl,cycloalkyl, cycloalkylalkyl, aryl, aralkyl, heteroaryl, heteroarylalkyl,heterocyclyl, or heterocyclylalkyl group.

In other embodiments, there are provided a second group of compounds ofFormula III:

stereoisomers thereof, tautomers thereof, solvates thereof, andpharmaceutically acceptable salts thereof; wherein

R₁ and R₂ are independently —H, —(CH₂)₀₋₆COOR′, —C(O)R″, or asubstituted or unsubstituted alkyl, cycloalkyl, cycloalkylalkyl,alkenyl, aryl, aralkyl, heteroaryl, heteroarylalkyl, heterocyclyl, orheterocyclylalkyl group; or R₁ and R₂ together are a methylene group;

R₃ and R₈ are independently —H, —OH, —Cl, —Br, —F, —I, —CN, —NH₂,—C(O)NH₂, —COOH, or a substituted or unsubstituted alkyl, alkenyl,alkoxy or aralkyl group;

R₃′ is —H, or R₃ and R₃′ together are an oxo group;

R₄ is —H, halogen, —OR′, —OSO₂R″, —OC(O)R″, —OC(O)OR″, —OC(O)NR′R″,—O-alkylene-NR′R′, —O-alkylene-OSO₂R″, —O-alkylene-S(O)₀₋₂R″,—O-alkylene-NR′SO₂R″, —O-alkylene-N(R′)C(O)R′, or a substituted orunsubstituted alkyl group;

R₅ and R₆ are independently —H, halogen, —OH, or a substituted orunsubstituted alkoxy group; or R₄ and R₅ together are a methylenedioxygroup, or R₅ and R₆ together are a methylenedioxy group;

R₇ is —H, halogen, —OH, or a substituted or unsubstituted alkyl oralkoxy group;

each R′ is independently a hydrogen, or a substituted or unsubstitutedalkyl, alkenyl, cycloalkyl, cycloalkylalkyl, aryl, aralkyl, heteroaryl,heteroarylalkyl, heterocyclyl, or heterocyclylalkyl group;

each R″ is independently a substituted or unsubstituted alkyl, alkenyl,cycloalkyl, cycloalkylalkyl, aryl, aralkyl, heteroaryl, heteroarylalkyl,heterocyclyl, or heterocyclylalkyl group;

with the proviso that when R₄ is —H, —OH or a C₁₋₄ alkoxy group, then R₅is not —H, —OH or a C₁₋₄ alkoxy group; and when R₁ and R₂ are both —CH₃or when R₁ and R₂ together are a methylene group, then R₅ is not OH or aC₁₋₂ alkoxy group, and R₄ and R₅ together are not a methylenedioxygroup; and when R₄ is OC(O)R″, then R₅ is not OC(O)R″ or methoxy.

In another aspect, a lipid lowering agent of the invention is part of apharmaceutical composition containing one or more excipients, carriers,or fillers. In one embodiment, the pharmaceutical composition ispackaged in unit dosage form. The unit dosage form is effective inlowering lipid levels (e.g., at least one of total cholesterol,LDL-cholesterol, triglyceride, and unesterified long chain fatty acids)in the bloodstream and/or in the liver when administered to a subject inneed thereof.

Still another aspect of the invention is a pharmaceutical pack or kitcontaining a lipid lowering agent according to the invention and asecond agent. The second agent can be a cholesterol uptake inhibitor, acholesterol synthesis inhibitor, a cholesterol absorption inhibitor, abile acid sequestrant, a vitamin, an antihypertensive agent, or aplatelet aggregation inhibitor. The second agent alternatively can be anHMG-CoA reductase inhibitor, an HMG-CoA synthase inhibitor, a squaleneepoxidase inhibitor, an acyl-CoA cholesterol acyltransferase (ACAT)inhibitor, a microsomal triglyceride transfer protein (MTP) inhibitor, aperoxisome proliferator-activated receptor (PPAR) agonist, or anAMP-activated protein kinase (AMPK) activator. The second agent can alsobe an agent that increases low density lipoprotein receptor (LDLR)expression. The second agent can be a berberine compound, such astetrahydroberberine.

Yet another aspect of the invention is a method of synthesizing14R-tetrahydropalmatine. The method includes treating berberine withboron trichloride in methylene chloride, methylating the product withmethyl iodide and potassium carbonate in dry acetone, and hydrogenatingthe product using an asymmetric hydrogenation catalyst to yield14R-tetrahydropalmatine.

DESCRIPTION OF THE DRAWINGS

FIG. 1 shows the determination of the stereochemical configuration ofCRDL by x-ray diffraction.

FIG. 2 shows the determination of the stereochemical configuration ofTHP by x-ray diffraction.

FIG. 3 shows the potent and dose-dependent effects of (+)-CLMD,14R-(+)-CRPM, 14R,13S-(+)-CRDL, and 14R-(+)-THP on LDLR mRNA expressionin HepG2 cells by a semi-quantitative RT-PCR analysis.

FIG. 4 shows the determination of the specific stereochemicalrequirements of +/−THP in the upregulation of LDLR mRNA expression.

FIG. 5 shows Western blot analysis of the activation of ERK in HepG2cells by 14R,13S-(+)-CRDL and 14R-(+)-THP.

FIG. 6 shows Western blot analysis of the induction of ACCphosphorylation by 14R-(+)-THP.

FIG. 7 shows a concentration vs. time curve of CRDL in hamstersfollowing intravenous administration of 2 mg/kg body weight. FIG. 7Ashows the results for individual animals, and FIG. 7B shows themean+/−standard deviation.

FIG. 8 shows a concentration vs. time curve of CRDL in hamstersfollowing oral administration of 20 mg/kg body weight. FIG. 8A shows theresults for individual animals, and FIG. 8B shows the mean+/−standarddeviation.

FIG. 9A is a bar graph showing plasma lipid levels in male Golden Syrianhamsters on a high cholesterol diet in control and d-THP (i.e.,(14R)-(+)-THP) treatment groups after 34 days of drug treatment by i.p.route. FIG. 9B shows weight gains in both groups.

FIG. 10A is a bar graph showing plasma lipid levels in male GoldenSyrian hamsters on a high cholesterol diet in control and (14R,13S)-CRDLtreated (10 mg/kg daily) groups after 17 days of drug treatment by i.p.route. FIG. 10B shows the elevated alanine aminotransaminase (ALT) andaspartate aminotransaminase (AST) in blood of the same hamsters incontrol and CRDL treatment group. CRDL treatment significantly reducedALT and AST in plasma. FIG. 10C shows that the blood levels of ureanitrogen (BUN) and glucose were not changed, while the level ofcreatinine (CREA) was reduced by 20% (p<0.001)

FIG. 11A shows a TC vs. time curve in Wister male rats treated with(14R,13S)-CRDL HCl and demonstrates that CRDL treatment lowered TC to33.8% compared to the control group and to 33.0% of the pretreatmentlevel. FIG. 11B shows a similar curve for LDL-c levels and shows thatthe LDL-c level was reduced by CRDL to 25.6% of control, and to 22.4% ofday 0 by CRDL treatment. FIG. 11C shows a similar curve for TG levelswhich indicates that the TG level was decreased to 29% of the controland to 27% of the pretreatment level (day 0). FIG. 11D is a bar graphshowing the serum levels of AST and ALT in Wister male rats treated with(14R,13S)-CRDL HCl and that of the control group and indicates thatliver function was not damaged by CRDL instead it was improved withstatistical significance. FIG. 11E is a similar bar graph which showsthe glucose, BUN and CREA levels and demonstrates that kidney functionand blood glucose level were not changed by the treatment with CRDL.

FIG. 12A shows the food intake vs. time curve for male Wister ratstreated with CRDL and the control group. After an initial decrease infood consumption during the first week, it increase and leveled off atstatistically similar level to the control group through the rest oftreatment times. FIG. 12B shows the change in body weight vs. time curefor the CRDL-treated Wister rats and the control group fed with high fatand high cholesterol diet and shows that, while the control group gainedover 30% of their body weight during the 4-weeks, the body weights ofWister rats in CRDL-treated group have maintained constant.

FIG. 13 shows that enantiomers of compounds disclosed herein withdextrorotary optical rotation elevate LDLR mRNA levels.

FIG. 14 is a Western blot showing that levels of phosphorylated andactivated ERK and AMPK were significantly increased in HepG2 cellstreated with compounds of the invention as compared to untreated controlcells.

FIG. 15 compares total cellular TG content of cells exposed to compoundsdisclosed herein.

FIG. 16 shows that the statin drug simvastatin increases LDLR mRNAlevels (top panel) but fails to lower cellular TG content (bottompanel).

FIG. 17 shows the combined effect of simvastatin and compounds of theinvention on LDLR mRNA levels.

FIG. 18 shows that compounds disclosed herein strongly inhibit the mRNAexpression of PCSK9.

FIG. 19A shows a LDLR mRNA level vs. concentration curve for compound 91and simvastatin and FIG. 19B shows a PCSK9 mRNA levels vs. concentrationcurve for curve compound 91.

DETAILED DESCRIPTION OF THE INVENTION

In various aspects, the present invention provides novel compounds,extracts, methods for reducing plasma and/or hepatic lipid levels, andmethods for treating hyperlipidemia, hypercholesterolemia,hypertriglyceridemia, hepatic steatosis and metabolic syndrome. Thecompounds provided herein can be formulated into pharmaceuticalcompositions and medicaments that are useful in the disclosed methods.Also provided are the use of the compounds and extracts in preparingpharmaceutical formulations and medicaments, the use of the compoundsand extracts in reducing plasma and/or hepatic lipid levels, and the useof the compounds and extracts in treating hyperlipidemia,hypercholesterolemia, hypertriglyceridemia, hepatic steatosis andmetabolic syndrome.

The following terms are used throughout as defined below.

Generally, reference to a certain element such as hydrogen or H is meantto include all isotopes of that element. For example, if an R group isdefined to include hydrogen or H, it also includes deuterium andtritium. Compounds comprising radioisotopes such as tritium, C¹⁴, P³²and S³⁵ are thus within the scope of the invention. Procedures forinserting such labels into the compounds of the invention will bereadily apparent to those skilled in the art based on the disclosureherein.

In general, “substituted” refers to an organic group as defined below(e.g., an alkyl group) in which one or more bonds to a hydrogen atomcontained therein are replaced by a bond to non-hydrogen or non-carbonatoms. Substituted groups also include groups in which one or more bondsto a carbon(s) or hydrogen(s) atom are replaced by one or more bonds,including double or triple bonds, to a heteroatom. Thus, a substitutedgroup is substituted with one or more substituents, unless otherwisespecified. In some embodiments, a substituted group is substituted with1, 2, 3, 4, 5, or 6 substituents. Examples of substituent groupsinclude: halogens (i.e., F, Cl, Br, and I); hydroxyls; alkoxy, alkenoxy,aryloxy, aralkyloxy, heterocyclyloxy, and heterocyclylalkoxy groups;carbonyls (oxo); carboxyls; esters; urethanes; oximes; hydroxylamines;alkoxyamines; aralkoxyamines; thiols; sulfides; sulfoxides; sulfones;sulfonyls; sulfonamides; amines; N-oxides; hydrazines; hydrazides;hydrazones; azides; amides; ureas; amidines; guanidines; enamines;imides; isocyanates; isothiocyanates; cyanates; thiocyanates; imines;nitro groups; nitriles (i.e., CN); and the like.

Substituted ring groups such as substituted cycloalkyl, aryl,heterocyclyl and heteroaryl groups also include rings and ring systemsin which a bond to a hydrogen atom is replaced with a bond to a carbonatom. Therefore, substituted cycloalkyl, aryl, heterocyclyl andheteroaryl groups may also be substituted with substituted orunsubstituted alkyl, alkenyl, and alkynyl groups as defined below.

Alkyl groups include straight chain and branched chain alkyl groupshaving from 1 to 12 carbon atoms, and typically from 1 to 10 carbons or,in some embodiments, from 1 to 8, 1 to 6, or 1 to 4 carbon atoms.Examples of straight chain alkyl groups include groups such as methyl,ethyl, n-propyl, n-butyl, n-pentyl, n-hexyl, n-heptyl, and n-octylgroups. Examples of branched alkyl groups include, but are not limitedto, isopropyl, iso-butyl, sec-butyl, tert-butyl, neopentyl, isopentyl,and 2,2-dimethylpropyl groups. Representative substituted alkyl groupsmay be substituted one or more times with substituents such as thoselisted above, and include without limitation haloalkyl (e.g.,trifluoromethyl), hydroxyalkyl, thioalkyl, aminoalkyl, alkylaminoalkyl,dialkylaminoalkyl, alkoxyalkyl, carboxyalkyl, and the like.

Cycloalkyl groups include mono-, bi- or tricyclic alkyl groups havingfrom 3 to 12 carbon atoms in the ring(s), or, in some embodiments, 3 to10, 3 to 8, or 3 to 4, 5, or 6 carbon atoms. Exemplary monocycliccycloalkyl groups include, but not limited to, cyclopropyl, cyclobutyl,cyclopentyl, cyclohexyl, cycloheptyl, and cyclooctyl groups. In someembodiments, the cycloalkyl group has 3 to 8 ring members, whereas inother embodiments the number of ring carbon atoms range from 3 to 5, 3to 6, or 3 to 7. Bi- and tricyclic ring systems include both bridgedcycloalkyl groups and fused rings, such as, but not limited to,bicyclo[2.1.1]hexane, adamantyl, decalinyl, and the like. Substitutedcycloalkyl groups may be substituted one or more times with,non-hydrogen and non-carbon groups as defined above. However,substituted cycloalkyl groups also include rings that are substitutedwith straight or branched chain alkyl groups as defined above.Representative substituted cycloalkyl groups may be mono-substituted orsubstituted more than once, such as, but not limited to, 2,2-, 2,3-,2,4- 2,5- or 2,6-disubstituted cyclohexyl groups, which may besubstituted with substituents such as those listed above.

Cycloalkylalkyl groups are alkyl groups as defined above in which ahydrogen or carbon bond of an alkyl group is replaced with a bond to acycloalkyl group as defined above. In some embodiments, cycloalkylalkylgroups have from 4 to 16 carbon atoms, 4 to 12 carbon atoms, andtypically 4 to 10 carbon atoms. Substituted cycloalkylalkyl groups maybe substituted at the alkyl, the cycloalkyl or both the alkyl andcycloalkyl portions of the group. Representative substitutedcycloalkylalkyl groups may be mono-substituted or substituted more thanonce, such as, but not limited to, mono-, di- or tri-substituted withsubstituents such as those listed above.

Alkenyl groups include straight and branched chain alkyl groups asdefined above, except that at least one double bond exists between twocarbon atoms. Alkenyl groups have from 2 to 12 carbon atoms, andtypically from 2 to 10 carbons or, in some embodiments, from 2 to 8, 2to 6, or 2 to 4 carbon atoms. In some embodiments, the alkenyl group hasone, two, or three carbon-carbon double bonds. Examples include, but arenot limited to vinyl, allyl, —CH═CH(CH₃), —CH═C(CH₃)₂, —C(CH₃)═CH₂,—C(CH₃)═CH(CH₃), —C(CH₂CH₃)═CH₂, among others. Representativesubstituted alkenyl groups may be mono-substituted or substituted morethan once, such as, but not limited to, mono-, di- or tri-substitutedwith substituents such as those listed above.

Cycloalkenyl groups include cycloalkyl groups as defined above, havingat least one double bond between two carbon atoms. In some embodimentsthe cycloalkenyl group may have one, two or three double bonds but doesnot include aromatic compounds. Cycloalkenyl groups have from 4 to 14carbon atoms, or, in some embodiments, 5 to 14 carbon atoms, 5 to 10carbon atoms, or even 5, 6, 7, or 8 carbon atoms. Examples ofcycloalkenyl groups include cyclohexenyl, cyclopentenyl,cyclohexadienyl, butadienyl, pentadienyl, and hexadienyl.

Cycloalkenylalkyl groups are alkyl groups as defined above in which ahydrogen or carbon bond of the alkyl group is replaced with a bond to acycloalkenyl group as defined above. Substituted cycloalkenylalkylgroups may be substituted at the alkyl, the cycloalkenyl or both thealkyl and cycloalkenyl portions of the group. Representative substitutedcycloalkenylalkyl groups may be substituted one or more times withsubstituents such as those listed above.

Alkynyl groups include straight and branched chain alkyl groups asdefined above, except that at least one triple bond exists between twocarbon atoms. Alkynyl groups have from 2 to 12 carbon atoms, andtypically from 2 to 10 carbons or, in some embodiments, from 2 to 8, 2to 6, or 2 to 4 carbon atoms. In some embodiments, the alkynyl group hasone, two, or three carbon-carbon triple bonds. Examples include, but arenot limited to —C≡CH, —C≡CH₃, —CH₂C≡CH₃, —C≡CH₂CH(CH₂CH₃)₂, amongothers. Representative substituted alkynyl groups may bemono-substituted or substituted more than once, such as, but not limitedto, mono-, di- or tri-substituted with substituents such as those listedabove.

Aryl groups are cyclic aromatic hydrocarbons that do not containheteroatoms. Aryl groups herein include monocyclic, bicyclic andtricyclic ring systems. Thus, aryl groups include, but are not limitedto, phenyl, azulenyl, heptalenyl, biphenyl, fluorenyl, phenanthrenyl,anthracenyl, indenyl, indanyl, pentalenyl, and naphthyl groups. In someembodiments, aryl groups contain 6-14 carbons, and in others from 6 to12 or even 6-10 carbon atoms in the ring portions of the groups. In someembodiments, the aryl groups are phenyl or naphthyl. Although the phrase“aryl groups” includes groups containing fused rings, such as fusedaromatic-aliphatic ring systems (e.g., indanyl, tetrahydronaphthyl, andthe like), it does not include aryl groups that have other groups, suchas alkyl or halo groups, bonded to one of the ring members. Rather,groups such as tolyl are referred to as substituted aryl groups.Representative substituted aryl groups may be mono-substituted orsubstituted more than once. For example, monosubstituted aryl groupsinclude, but are not limited to, 2-, 3-, 4-, 5-, or 6-substituted phenylor naphthyl groups, which may be substituted with substituents such asthose listed above.

Aralkyl groups are alkyl groups as defined above in which a hydrogen orcarbon bond of an alkyl group is replaced with a bond to an aryl groupas defined above. In some embodiments, aralkyl groups contain 7 to 16carbon atoms, 7 to 14 carbon atoms, or 7 to 10 carbon atoms. Substitutedaralkyl groups may be substituted at the alkyl, the aryl or both thealkyl and aryl portions of the group. Representative aralkyl groupsinclude but are not limited to benzyl and phenethyl groups and fused(cycloalkylaryl)alkyl groups such as 4-indanylethyl. Representativesubstituted aralkyl groups may be substituted one or more times withsubstituents such as those listed above.

Heterocyclyl groups include aromatic (also referred to as heteroaryl)and non-aromatic ring compounds containing 3 or more ring members, ofwhich one or more is a heteroatom such as, but not limited to, N, O, andS. In some embodiments, the heterocyclyl group contains 1, 2, 3 or 4heteroatoms. In some embodiments, heterocyclyl groups include mono-, bi-and tricyclic rings having 3 to 16 ring members, whereas other suchgroups have 3 to 6, 3 to 10, 3 to 12, or 3 to 14 ring members.Heterocyclyl groups encompass aromatic, partially unsaturated andsaturated ring systems, such as, for example, imidazolyl, imidazolinyland imidazolidinyl groups. The phrase “heterocyclyl group” includesfused ring species including those comprising fused aromatic andnon-aromatic groups, such as, for example, benzotriazolyl,2,3-dihydrobenzo[1,4]dioxinyl, and benzo[1,3]dioxolyl. The phrase alsoincludes bridged polycyclic ring systems containing a heteroatom suchas, but not limited to, quinuclidyl. However, the phrase does notinclude heterocyclyl groups that have other groups, such as alkyl, oxoor halo groups, bonded to one of the ring members. Rather, these arereferred to as “substituted heterocyclyl groups”. Heterocyclyl groupsinclude, but are not limited to, aziridinyl, azetidinyl, pyrrolidinyl,imidazolidinyl, pyrazolidinyl, thiazolidinyl, tetrahydrothiophenyl,tetrahydrofuranyl, dioxolyl, furanyl, thiophenyl, pyrrolyl, pyrrolinyl,imidazolyl, imidazolinyl, pyrazolyl, pyrazolinyl, triazolyl, tetrazolyl,oxazolyl, isoxazolyl, thiazolyl, thiazolinyl, isothiazolyl,thiadiazolyl, oxadiazolyl, piperidyl, piperazinyl, morpholinyl,thiomorpholinyl, tetrahydropyranyl, tetrahydrothiopyranyl, oxathiane,dioxyl, dithianyl, pyranyl, pyridyl, pyrimidinyl, pyridazinyl,pyrazinyl, triazinyl, dihydropyridyl, dihydrodithiinyl,dihydrodithionyl, homopiperazinyl, quinuclidyl, indolyl, indolinyl,isoindolyl, azaindolyl (pyrrolopyridyl), indazolyl, indolizinyl,benzotriazolyl, benzimidazolyl, benzofuranyl, benzothiophenyl,benzthiazolyl, benzoxadiazolyl, benzoxazinyl, benzodithiinyl,benzoxathiinyl, benzothiazinyl, benzoxazolyl, benzothiazolyl,benzothiadiazolyl, benzo[1,3]dioxolyl, pyrazolopyridyl, imidazopyridyl(azabenzimidazolyl), triazolopyridyl, isoxazolopyridyl, purinyl,xanthinyl, adeninyl, guaninyl, quinolinyl, isoquinolinyl, quinolizinyl,quinoxalinyl, quinazolinyl, cinnolinyl, phthalazinyl, naphthyridinyl,pteridinyl, thianaphthyl, dihydrobenzothiazinyl, dihydrobenzofuranyl,dihydroindolyl, dihydrobenzodioxinyl, tetrahydroindolyl,tetrahydroindazolyl, tetrahydrobenzimidazolyl, tetrahydrobenzotriazolyl,tetrahydropyrrolopyridyl, tetrahydropyrazolopyridyl,tetrahydroimidazopyridyl, tetrahydrotriazolopyridyl, andtetrahydroquinolinyl groups. Representative substituted heterocyclylgroups may be mono-substituted or substituted more than once, such as,but not limited to, pyridyl or morpholinyl groups, which are 2-, 3-, 4-,5-, or 6-substituted, or disubstituted with various substituents such asthose listed above.

Heteroaryl groups are aromatic ring compounds containing 5 or more ringmembers, of which, one or more is a heteroatom such as, but not limitedto, N, O, and S. Heteroaryl groups include, but are not limited to,groups such as pyrrolyl, pyrazolyl, triazolyl, tetrazolyl, oxazolyl,isoxazolyl, thiazolyl, pyridinyl, pyridazinyl, pyrimidinyl, pyrazinyl,thiophenyl, benzothiophenyl, furanyl, benzofuranyl, indolyl, azaindolyl(pyrrolopyridinyl), indazolyl, benzimidazolyl, imidazopyridinyl(azabenzimidazolyl), pyrazolopyridinyl, triazolopyridinyl,benzotriazolyl, benzoxazolyl, benzothiazolyl, benzothiadiazolyl,imidazopyridinyl, isoxazolopyridinyl, thianaphthyl, purinyl, xanthinyl,adeninyl, guaninyl, quinolinyl, isoquinolinyl, tetrahydroquinolinyl,quinoxalinyl, and quinazolinyl groups. Heteroaryl groups include fusedring compounds in which all rings are aromatic such as indolyl groupsand include fused ring compounds in which only one of the rings isaromatic, such as 2,3-dihydro indolyl groups. Although the phrase“heteroaryl groups” includes fused ring compounds, the phrase does notinclude heteroaryl groups that have other groups bonded to one of thering members, such as alkyl groups. Rather, heteroaryl groups with suchsubstitution are referred to as “substituted heteroaryl groups.”Representative substituted heteroaryl groups may be substituted one ormore times with various substituents such as those listed above.

Heterocyclylalkyl groups are alkyl groups as defined above in which ahydrogen or carbon bond of an alkyl group is replaced with a bond to aheterocyclyl group as defined above. Substituted heterocyclylalkylgroups may be substituted at the alkyl, the heterocyclyl or both thealkyl and heterocyclyl portions of the group. Representativeheterocyclyl alkyl groups include, but are not limited to,morpholin-4-yl-ethyl, furan-2-yl-methyl, imidazol-4-yl-methyl,pyridin-3-yl-methyl, tetrahydrofuran-2-yl-ethyl, and indol-2-yl-propyl.Representative substituted heterocyclylalkyl groups may be substitutedone or more times with substituents such as those listed above.

Heteroaralkyl groups are alkyl groups as defined above in which ahydrogen or carbon bond of an alkyl group is replaced with a bond to aheteroaryl group as defined above. Substituted heteroaralkyl groups maybe substituted at the alkyl, the heteroaryl or both the alkyl andheteroaryl portions of the group. Representative substitutedheteroaralkyl groups may be substituted one or more times withsubstituents such as those listed above.

Groups described herein having two or more points of attachment (i.e.,divalent, trivalent, or polyvalent) within the compound of the inventionare designated by use of the suffix, “ene.” For example, divalent alkylgroups are alkylene groups, divalent aryl groups are arylene groups,divalent heteroaryl groups are divalent heteroarylene groups, and soforth. Substituted groups having a single point of attachment to thecompound of the invention are not referred to using the “ene”designation. Thus, e.g., chloroethyl is not referred to herein aschloroethylene.

Alkoxy groups are hydroxyl groups (—OH) in which the bond to thehydrogen atom is replaced by a bond to a carbon atom of a substituted orunsubstituted alkyl group as defined above. Examples of linear alkoxygroups include but are not limited to methoxy, ethoxy, propoxy, butoxy,pentoxy, hexoxy, and the like. Examples of branched alkoxy groupsinclude but are not limited to isopropoxy, sec-butoxy, tert-butoxy,isopentoxy, isohexoxy, and the like. Examples of cycloalkoxy groupsinclude but are not limited to cyclopropyloxy, cyclobutyloxy,cyclopentyloxy, cyclohexyloxy, and the like. Representative substitutedalkoxy groups may be substituted one or more times with substituentssuch as those listed above.

The terms “alkanoyl” and “alkanoyloxy” as used herein can refer,respectively, to —C(O)-alkyl groups and —O—C(O)-alkyl groups, eachcontaining 2-5 carbon atoms.

The terms “aryloxy” and “arylalkoxy” refer to, respectively, asubstituted or unsubstituted aryl group bonded to an oxygen atom and asubstituted or unsubstituted aralkyl group bonded to the oxygen atom atthe alkyl. Examples include but are not limited to phenoxy, naphthyloxy,and benzyloxy. Representative substituted aryloxy and arylalkoxy groupsmay be substituted one or more times with substituents such as thoselisted above.

The term “carboxylate” as used herein refers to a —COOH group.

The term “ester” as used herein refers to —COOR³⁰ groups. R³⁰ is asubstituted or unsubstituted alkyl, cycloalkyl, alkenyl, alkynyl, aryl,aralkyl, heterocyclylalkyl or heterocyclyl group as defined herein.

The term “amide” (or “amido”) includes C- and N-amide groups, i.e.,—C(O)NR³¹R³², and —NR³¹C(O)R³² groups, respectively. R³¹ and R³² areindependently hydrogen, or a substituted or unsubstituted alkyl,alkenyl, alkynyl, cycloalkyl, aryl, aralkyl, heterocyclylalkyl orheterocyclyl group as defined herein. Amido groups therefore include butare not limited to carbamoyl groups (—C(O)NH₂) and formamide groups(—NHC(O)H). In some embodiments, the amide is —NR³¹C(O)—(C₁₋₅ alkyl) andthe group is termed “carbonylamino,” and in others the amide is—NHC(O)-alkyl and the group is termed “alkanoylamino.”

The term “nitrile” or “cyano” as used herein refers to the —CN group.

Urethane groups include N- and O-urethane groups, i.e., —NR³³C(O)OR³⁴and —OC(O)NR³³R³⁴ groups, respectively. R³³ and R³⁴ are independently asubstituted or unsubstituted alkyl, alkenyl, alkynyl, cycloalkyl, aryl,aralkyl, heterocyclylalkyl, or heterocyclyl group as defined herein. R³³may also be H.

The term “amine” (or “amino”) as used herein refers to —NR³⁵R³⁶ groups,wherein R³⁵ and R³⁶ are independently hydrogen, or a substituted orunsubstituted alkyl, alkenyl, alkynyl, cycloalkyl, aryl, aralkyl,heterocyclylalkyl or heterocyclyl group as defined herein. In someembodiments, the amine is alkylamino, dialkylamino, arylamino, oralkylarylamino. In other embodiments, the amine is NH₂, methylamino,dimethylamino, ethylamino, diethylamino, propylamino, isopropylamino,phenylamino, or benzylamino.

The term “sulfonamido” includes S- and N-sulfonamide groups, i.e.,—SO₂NR³⁸R³⁹ and —NR³⁸SO₂R³⁹ groups, respectively. R³⁸ and R³⁹ areindependently hydrogen, or a substituted or unsubstituted alkyl,alkenyl, alkynyl, cycloalkyl, aryl, aralkyl, heterocyclylalkyl, orheterocyclyl group as defined herein. Sulfonamido groups thereforeinclude but are not limited to sulfamoyl groups (—SO₂NH₂). In someembodiments herein, the sulfonamido is —NHSO₂-alkyl and is referred toas the “alkylsulfonylamino” group.

The term “thiol” refers to —SH groups, while sulfides include —SR⁴⁰groups, sulfoxides include —S(O)R⁴¹ groups, sulfones include —SO₂R⁴²groups, and sulfonyls include —SO₂OR⁴³. R⁴⁰, R⁴¹, R⁴², and R⁴³ are eachindependently a substituted or unsubstituted alkyl, cycloalkyl, alkenyl,alkynyl, aryl aralkyl, heterocyclyl or heterocyclylalkyl group asdefined herein. In some embodiments the sulfide is an alkylthio group,—S-alkyl.

The term “urea” refers to —NR⁴⁴—C(O)—NR⁴⁵R⁴⁶ groups. R⁴⁴, R⁴⁵, and R⁴⁶groups are independently hydrogen, or a substituted or unsubstitutedalkyl, alkenyl, alkynyl, cycloalkyl, aryl, aralkyl, heterocyclyl, orheterocyclylalkyl group as defined herein.

The term “amidine” refers to —C(NR⁴⁷)NR⁴⁸R⁴⁹ and —NR⁴⁷C(NR⁴⁸)R⁴⁹,wherein R⁴⁷, R⁴⁸, and R⁴⁹ are each independently hydrogen, or asubstituted or unsubstituted alkyl, cycloalkyl, alkenyl, alkynyl, arylaralkyl, heterocyclyl or heterocyclylalkyl group as defined herein.

The term “guanidine” refers to —NR C(NR⁵¹)NR⁵²R⁵³, wherein R⁵⁰, R⁵¹, R⁵²and R⁵³ are each independently hydrogen, or a substituted orunsubstituted alkyl, cycloalkyl, alkenyl, alkynyl, aryl aralkyl,heterocyclyl or heterocyclylalkyl group as defined herein.

The term “enamine” refers to —C(R⁵⁴)═C(R⁵⁵)NR⁵⁶R⁵⁷ and—NR⁵⁴C(R⁵⁵)═C(R⁵⁶)R⁵⁷, wherein R⁵⁴, R⁵⁵, R⁵⁶ and R⁵⁷ are eachindependently hydrogen, a substituted or unsubstituted alkyl,cycloalkyl, alkenyl, alkynyl, aryl aralkyl, heterocyclyl orheterocyclylalkyl group as defined herein.

The term “halogen” or “halo” as used herein refers to bromine, chlorine,fluorine, or iodine. In some embodiments, the halogen is fluorine. Inother embodiments, the halogen is chlorine or bromine.

The term “hydroxy” as used herein can refer to —OH or its ionized form,—O⁻.

The term “imide” refers to —C(O)NR⁵⁸C(O)R⁵⁹, wherein R⁵⁸ and R⁵⁹ areeach independently hydrogen, or a substituted or unsubstituted alkyl,cycloalkyl, alkenyl, alkynyl, aryl aralkyl, heterocyclyl orheterocyclylalkyl group as defined herein.

The term “imine” refers to —CR⁶⁰(NR⁶¹) and —N(CR⁶⁰R⁶¹) groups, whereinR⁶⁰ and R⁶¹ are each independently hydrogen or a substituted orunsubstituted alkyl, cycloalkyl, alkenyl, alkynyl, aryl aralkyl,heterocyclyl or heterocyclylalkyl group as defined herein, with theproviso that R⁶⁰ and R⁶¹ are not both simultaneously hydrogen.

The term “nitro” as used herein refers to an —NO₂ group.

The term “trifluoromethyl” as used herein refers to —CF₃.

The term “trifluoromethoxy” as used herein refers to —OCF₃.

As will be understood by one skilled in the art, for any and allpurposes, particularly in terms of providing a written description, allranges disclosed herein also encompass any and all possible subrangesand combinations of subranges thereof. Any listed range can be easilyrecognized as sufficiently describing and enabling the same range beingbroken down into at least equal halves, thirds, quarters, fifths,tenths, etc. As a non-limiting example, each range discussed herein canbe readily broken down into a lower third, middle third and upper third,etc. As will also be understood by one skilled in the art all languagesuch as “up to,” “at least,” “greater than,” “less than,” and the likeinclude the number recited and refer to ranges which can be subsequentlybroken down into subranges as discussed above. Finally, as will beunderstood by one skilled in the art, a range includes each individualmember. Thus, for example, a group having 1-3 atoms refers to groupshaving 1, 2, or 3 atoms. Similarly, a group having 1-5 atoms refers togroups having 1, 2, 3, 4, or 5 atoms, and so forth.

Pharmaceutically acceptable salts of compounds described herein arewithin the scope of the present invention and include acid or baseaddition salts which retain the desired pharmacological activity and isnot biologically undesirable (e.g., the salt is not unduly toxic,allergenic, or irritating, and is bioavailable). When the compound ofthe invention has a basic group, such as, for example, an amino group,pharmaceutically acceptable salts can be formed with inorganic acids(such as hydrochloric acid, hydroboric acid, nitric acid, sulfuric acid,and phosphoric acid), organic acids (e.g. alginate, formic acid, aceticacid, benzoic acid, gluconic acid, fumaric acid, oxalic acid, tartaricacid, lactic acid, maleic acid, citric acid, succinic acid, malic acid,methanesulfonic acid, benzenesulfonic acid, naphthalene sulfonic acid,and p-toluenesulfonic acid) or acidic amino acids (such as aspartic acidand glutamic acid). When the compound of the invention has an acidicgroup, such as for example, a carboxylic acid group, it can form saltswith metals, such as alkali and earth alkali metals (e.g. Na⁺, Li⁺, K⁺,Ca²⁺, Mg²⁺, Zn²⁺), ammonia or organic amines (e.g. dicyclohexylamine,trimethylamine, triethylamine, pyridine, picoline, ethanolamine,diethanolamine, triethanolamine) or basic amino acids (e.g. arginine,lysine and ornithine). Such salts can be prepared in situ duringisolation and purification of the compounds or by separately reactingthe purified compound in its free base or free acid form with a suitableacid or base, respectively, and isolating the salt thus formed.

Those of skill in the art will appreciate that compounds of theinvention may exhibit the phenomena of tautomerism, conformationalisomerism, geometric isomerism and/or stereoisomerism. As the formuladrawings within the specification and claims can represent only one ofthe possible tautomeric, conformational isomeric, stereochemical orgeometric isomeric forms, it should be understood that the inventionencompasses any tautomeric, conformational isomeric, stereochemicaland/or geometric isomeric forms of the compounds having one or more ofthe utilities described herein, as well as mixtures of these variousdifferent forms.

“Tautomers” refers to isomeric forms of a compound that are inequilibrium with each other. The presence and concentrations of theisomeric forms will depend on the environment the compound is found inand may be different depending upon, for example, whether the compoundis a solid or is in an organic or aqueous solution. For example, inaqueous solution, imidazoles may exhibit the following isomeric forms,which are referred to as tautomers of each other:

As readily understood by one skilled in the art, a wide variety offunctional groups and other structures may exhibit tautomerism, and alltautomers of compounds as described herein are within the scope of thepresent invention.

Stereoisomers of compounds (also known as optical isomers) include allchiral, diastereomeric, and racemic forms of a structure, unless thespecific stereochemistry is expressly indicated. Thus, compounds used inthe present invention include enriched or resolved optical isomers atany or all asymmetric atoms as are apparent from the depictions. Bothracemic and diastereomeric mixtures, as well as the individual opticalisomers can be isolated or synthesized so as to be substantially free oftheir enantiomeric or diastereomeric partners, and these stereoisomersare all within the scope of the invention.

The compounds of the invention may exist as solvates, especiallyhydrates. Hydrates may form during manufacture of the compounds orcompositions comprising the compounds, or hydrates may form over timedue to the hygroscopic nature of the compounds. Compounds of theinvention may exist as organic solvates as well, including DMF, ether,and alcohol solvates among others. The identification and preparation ofany particular solvate is within the skill of the ordinary artisan ofsynthetic organic or medicinal chemistry.

Lipids include synthetic or naturally-occurring fat-soluble compounds,and include both neutral and amphipathic molecules. Amphipathic lipidstypically comprise a hydrophilic component and a hydrophobic component.Exemplary lipids include fatty acids, triglycerides, neutral fats,phosphatides, glycolipids, aliphatic alcohols, waxes, terpenes, steroidssuch as cholesterol, and surfactants.

A “lipid lowering agent” as used herein refers to compounds and plantextracts containing compounds that have one or more of the followingeffects when administered to a subject: increasing the hepaticexpression of LDLR; increasing the half-life of LDLR mRNA inhepatocytes; increasing hepatic uptake of plasma LDL, cholesterol, ortriglycerides; enhancing hepatic fatty acid oxidation, reducing hepatictriglyceride synthesis and secretion, and reducing the plasma and/orhepatic levels of total cholesterol, LDL-cholesterol, VLDL-cholesterol,or triglycerides. Lipid lowering agents as disclosed herein includecompounds of Formulas I, II, III, and IV as well as extracts of plants(e.g., a Corydalis extract) containing one or more of such compounds,related compounds, or other alkaloid compounds having lipid loweringactivity.

A “Corydalis extract” as used herein refers to a mixture of chemicalcompounds obtained from any portion of plant tissue obtained from one ormore species of plant from the genus Corydalis. Such an extract can beobtained, for example, by grinding or homogenizing leaves, shoots, orroots from a Corydalis plant in a suitable solvent, e.g., an aqueousbuffer, an organic solvent, or a mixture thereof. A Corydalis extractcan be further processed by centrifugation, filtration, drying, furtherextraction, partitioning, chromatographic separation, or other chemicalor biochemical processing steps. Furthermore, a Corydalis extract can beformulated for administration to a subject by the addition of one ormore carriers, diluents, salts, buffers, flavoring agents,pharmaceutical agents, or nutrients. Extracts of lipid lowering agentsfrom other alkaloid containing plants maybe prepared in a similarfashion. In certain embodiments, a Corydalis extract according to theinvention does not contain a 14-reductase inhibitor.

A “compound” or “derivative” as used herein refers to a chemicalcompound, either in partially purified or substantially pure form, whicheither has been obtained from a plant extract, such as a Corydalisextract, by one or more purification steps or which has been produced bychemical synthesis from any desired starting materials. A compound orderivative according to the invention can be used either as a racemicmixture or as a pure stereoisomer. Preferred are pure stereoisomerswhich have activity as a lipid lowering agent.

A “partially purified” compound or derivative as used herein refers to aCorydalis compound or derivative thereof which is present in a chemicalmixture that has been subjected to at least one separation orpurification step resulting in the removal of at least one otherchemical substance originally present in the initial extract orsynthetic mixture containing the compound or derivative. A“substantially pure” compound or derivative is one which has beenseparated or purified to render the compound or derivative as the majorchemical component of the substantially pure compound or derivative,i.e., comprising at least 50%, or in some embodiments at least 70%, atleast 90%, or at least 95% or 99% on a molar basis.

In one aspect, the present invention provides methods of reducing plasmaand/or hepatic lipid levels in a subject in need thereof, whichcomprises administering to said subject a lipid-lowering effectiveamount of a compound, extract or composition as described herein. Thelipid level to be reduced can be one or more of total cholesterol,LDL-cholesterol (LDL-c), triglycerides (TG), and unesterified long chainfatty acids.

The compounds, extracts and compositions described herein may be used inthe treatment or prophylaxis of diseases that include, for example,hyperlipidemia, hypercholesterolemia, hypertriglyceridemia, fatty liver(hepatic steatosis), and metabolic syndrome. Methods of treatmentinclude administering to a subject in need thereof a therapeuticallyeffective amount of a compound, composition or extract described herein.The compounds of the invention can also be used in the treatment orprophylaxis of a disease state or malady characterized by or associatedwith elevated plasma or hepatic cholesterol or triglycerides. Generally,prophylactic or prophylaxis relates to a reduction in the likelihood ofthe patient developing a disorder such as hyperlipidemia,hypercholesterolemia, hypertriglyceridemia, fatty liver, or metabolicsyndrome or proceeding to a diagnosis state for the disorder. Forexample, the compounds of the invention can be used prophylacticly as ameasure designed to preserve health and prevent the spread or maturationof disease in a patient. It is also appreciated that the various modesof treatment or prevention of a disease such as hyperlipidemia,hypercholesterolemia, hypertriglyceridemia, fatty liver, or metabolicsyndrome can mean “substantial” treatment or prevention, which includestotal but also less than total treatment or prevention, and in whichsome biologically or medically relevant result is achieved. Furthermore,treatment or treating as well as alleviating can refer to therapeutictreatment and prophylactic or preventative measures in which the objectis to prevent, slow down (lessen) a disease state, condition or malady.For example, a subject can be successfully treated forhypercholesterolemia if, after receiving through administration aneffective or therapeutic amount of one or more compounds describedherein, the subject shows observable and/or measurable reduction in orabsence of one or more signs and symptoms of the particular disease suchas, but not limited to, reduced plasma total cholesterol, reduced plasmaLDL-cholesterol, increased hepatic expression of LDL receptor (LDLR),reduced plasma triglycerides, reduced morbidity and mortality, orimprovement in quality of life issues. The invention also provides formethods of administering one or more compounds of the invention to apatient in an effective amount for the treatment or prophylaxis of adisease such as, for example, hyperlipidemia, hypercholesterolemia,hypertriglyceridemia, fatty liver, or metabolic syndrome.

While not wishing to be bound by theory, it is believed that thecompounds, extracts and compositions disclosed herein reduce lipidlevels by increasing the hepatic expression of LDLR by increasing thestability of LDLR mRNA, by increasing LDLR gene transcription, byinhibiting the degradation of LDLR protein mediated through theproprotein convertase subtilisin/kexin type 9 (PCSK9), or all of theabove potential cellular mechanisms. Increasing LDLR levels in the liverincreases the uptake and processing of plasma LDL-c, resulting inreduced plasma levels of cholesterol, LDL-c, and triglycerides. Inaddition, the compounds may increase phosphorylation of acetyl CoAcarboxylase (ACC) via the activation of AMP-activated protein likase(AMPK). Increased phosphorylation of ACC enhances fatty acid oxidationin the liver, leading to reduced hepatic TG accumulation and secretionof TG in the form of VLDL, which also contributes to the decreasedplasma levels of TG, LDL-c, total cholesterol, and unesterified longchain fatty acids, resulting in the prevention or treatment of diseasesrelated to hyperlipidemia.

Hence, in another aspect, the invention provides methods of increasingLDLR expression, comprising administering to a subject in need thereof atherapeutically effective amount of a compound, extract or compositionas described herein, whereby LDLR expression in said subject isincreased. In another aspect of the invention, there are providedmethods of decreasing plasma LDL-cholesterol and/or plasmatriglycerides, comprising administering to a subject in need thereof atherapeutically effective amount of a compound, extract or compositionas described herein, whereby plasma LDL-cholesterol in said subject isdecreased.

“Effective amount” refers to the amount of a compound, extract orcomposition required to produce a desired effect. One example of aneffective amount includes amounts or dosages that yield acceptabletoxicity and bioavailability levels for therapeutic (pharmaceutical) useincluding, but not limited to, the treatment or prophylaxis ofhyperlipidemia, hypercholesterolemia, hypertriglyceridemia, fatty liver,or metabolic syndrome. Another example of an effective amount includesamounts or dosages that are capable of preventing elevated plasma orhepatic cholesterol or triglycerides.

As used herein, a “subject” or “patient” is a mammal, such as a cat,dog, rodent or primate. Typically the subject is a human, and,preferably, a human suspected of having a disease associated withelevated plasma or hepatic cholesterol or triglycerides such ashyperlipidemia, hypercholesterolemia, hypertriglyceridemia, fatty liver,or metabolic syndrome. Subjects may further include mammals withelevated LDL levels, elevated VLDL levels, or diseases aggravated ortriggered by hyperlipidemia such as cardiovascular diseases, including,atherosclerosis, coronary artery disease, angina pectoris, carotidartery disease, strokes, cerebral arteriosclerosis, myocardialinfarction, cerebral infarction, restenosis following balloonangioplasty, intermittent claudication, high blood pressure,dyslipidemia post-prandial lipidemia and xanthoma. The term “subject”and “patient” can be used interchangeably.

In another aspect, the invention provides lipid lowering agents,including compounds, extracts and compositions thereof. The compounds,extracts and compositions may be used in the lipid lowering methods andtreatments described herein. In one embodiment, the invention provides acompound of Formula I, a compound of Formula II, stereoisomers thereof,tautomers thereof, solvates thereof, and/or pharmaceutically acceptablesalt thereof,

For compounds of either Formula I or Formula II, R₁, R₂, R₃, R₄, R₅, andR₆ are selected (independently, collectively, or in any combination)from H, halogen, hydroxy, C₁-C₆ alkyl, alkoxy, nitro, amino, aminoalkyl,trifluoromethyl, trifluoromethoxy, cycloalkyl, alkanoyl, alkanoyloxy,nitrile, dialkylamino, alkenyl, hydroxyalkyl, alkylaminoalkyl,aminoalkyl, dialkylaminoalkyl, haloalkyl, carboxyalkyl, alkoxyalkyl,carboxy, alkanoylamino, carbonylamino, carbamoyl, alkylsulfonylamino,and heterocyclo groups. Preferably, R₁, R₂, R₃, R₄, R₅, and R₆ are nothalogen when halogen would be covalently bonded to oxygen. In oneaspect, the compounds of the invention can also comprise one or morehalogens as substituents at any position of Formula I or Formula II. Insome embodiments, compounds of Formula I have the 14R-(+) stereochemicalconfiguration and compounds of Formula II have the 1R-(+) stereochemicalconfiguration.

In another embodiment, there are provided a first group of compounds ofFormula III and compounds of Formula IV, as well as stereoisomersthereof, tautomers thereof, solvates thereof, and pharmaceuticallyacceptable salt thereof.

In compounds of Formula III and IV,

R₁ and R₂ are independently —H, —(CH₂)₀₋₆COOR′, —C(O)R″, or asubstituted or unsubstituted alkyl, cycloalkyl, cycloalkylalkyl,alkenyl, aryl, aralkyl, heteroaryl, heteroarylalkyl, heterocyclyl, orheterocyclylalkyl group; or R₁ and R₂ together are a methylene group;

R₃ and R₈ are independently —H, —OH, —Cl, —Br, —F, —I, —CN, —NH₂,—C(O)NH₂, —COOH, or a substituted or unsubstituted alkyl, alkoxy,alkenyl, or aralkyl group;

R₃′ is —H, or R₃ and R₃′ together are an oxo group;

R₄ is —H, halogen, —OR′, —OSO₂R″, —OC(O)R″, —OC(O)OR″, —OC(O)NR′R″,—O-alkylene-NR′R′, —O-alkylene-OSO₂R″, —O-alkylene-S(O)₀₋₂R″,—O-alkylene-NR′SO₂R″, —O-alkylene-N(R′)C(O)R′, or a substituted orunsubstituted alkyl group;

R₅ and R₆ are independently —H, halogen, —OH, or a substituted orunsubstituted alkoxy group; or R₄ and R₅ together are a methylenedioxygroup, or R₅ and R₆ together are a methylenedioxy group;

R₇ is —H, halogen, —OH, or a substituted or unsubstituted alkyl oralkoxy group;

R₉ is —H or a substituted or unsubstituted alkyl group;

each R′ is independently a hydrogen, or a substituted or unsubstitutedalkyl, alkenyl, cycloalkyl, cycloalkylalkyl, aryl, aralkyl, heteroaryl,heteroarylalkyl, heterocyclyl, or heterocyclylalkyl group;

each R″ is independently a substituted or unsubstituted alkyl, alkenyl,cycloalkyl, cycloalkylalkyl, aryl, aralkyl, heteroaryl, heteroarylalkyl,heterocyclyl, or heterocyclylalkyl group.

In some embodiments of the first group of compounds of Formula III andcompounds of Formula IV,

R₁ and R₂ are independently —H, —(CH₂)₀₋₆COOR′, —C(O)R″, or asubstituted or unsubstituted alkyl, cycloalkyl, cycloalkylalkyl,alkenyl, aryl, aralkyl, heteroaryl, heteroarylalkyl, heterocyclyl, orheterocyclylalkyl group; or R₁ and R₂ together are a methylene group;

R₃ and R₈ are independently —H, —OH, —Cl, —Br, —F, —I, —CN, —NH₂,—C(O)NH₂, —COOH, or a substituted or unsubstituted alkyl, alkenyl,alkoxy, or aralkyl group;

R₃′ is —H, or R₃ and R₃′ together are an oxo group;

R₄ is —H, —OR′, —OSO₂R″, —OC(O)R″, —OC(O)OR″, —OC(O)NR′R″,—O-alkylene-NR′R′, —O-alkylene-OSO₂R″, —O-alkylenelkylene-NR′SO₂R″,—O-alkylene-N(R′)C(O)R′, or a substituted or unsubstituted alkyl group;

R₅ and R₆ are independently —H, halogen, —OH, or a substituted orunsubstituted alkoxy group; or R₄ and R₅ together are a methylenedioxygroup, or R₅ and R₆ together are a methylenedioxy group;

R⁷ is —H, —Br, —Cl, or —F;

each R′ is independently a hydrogen, or a substituted or unsubstitutedalkyl, alkenyl, cycloalkyl, cycloalkylalkyl, aryl, aralkyl, heteroaryl,heteroarylalkyl, heterocyclyl, or heterocyclylalkyl group;

each R″ is independently a substituted or unsubstituted alkyl, alkenyl,cycloalkyl, cycloalkylalkyl, aryl, aralkyl, heteroaryl, heteroarylalkyl,heterocyclyl, or heterocyclylalkyl group.

In other embodiments of the first group of compounds of Formula III andcompounds of Formula IV,

R₁ and R₂ are independently —H, —(CH₂)₀₋₂COOR′, —C(O)(CH₂)₀₋₂R″, or aunsubstituted C₁₋₆ alkyl group; or R₁ and R₂ together are a methylenegroup;

R₃ and R₃′ are each —H, or R₃ and R₃′ together are an oxo group;

R₄ is —H, —OH, or a substituted or unsubstituted C₁₋₆ alkoxy, C₇₋₁₄aralkoxy, —OC(O)—(C₁₋₆ alkyl), —OC(O)-(aryl), —OC(O)O-(aryl),—OC(O)—NH-(aryl), —O—(C₂₋₆ alkylene)-NH—(C₂₋₆ alkyl), —O—(C₂₋₆alkylene)-NH-(tetrahydropyran), —O—(C₂₋₆ alkylene)-NH-(thiomorpholinedioxide), —O—(C₂₋₆ alkylene)-NH-(piperidinyl), —O—(C₂₋₆alkylene)-NH-(piperazinyl), —O—(C₂₋₆ alkylene)-NH-(morpholinyl),—O—(C₂₋₆ alkylene)-NH-(aralkyl), —O—(C₂₋₆ alkylene)-NH-(cyclopropyl),—OSO₂—(C₃₋₆ cycloalkyl), —OSO₂-(aryl), —O—(C₂₋₆ alkylene)-OSO₂-(aryl),—OSO₂-(aralkyl), —O—(C₂₋₆ alkylene)-OSO₂-(heteroaryl), —OSO₂—(C₁₋₆alkyl), —OSO₂-(pyridyl), —OSO₂(thiazolyl), —O—(C₂₋₆alkylene)-NHSO₂-(aryl), —O—(C₂₋₆ alkylene)-NHSO₂-(heteroaryl), —O—(C₂₋₆alkylene)-NHC(O)-(aryl), —O—(C₂₋₆ alkylene)-NHC(O)-(heteroaryl),—O—(C₀₋₄ alkyl)pyridyl, —O—(C₀₋₄ alkyl)pyrimidinyl, —O—(C₀₋₄alkyl)morpholinyl, —O—(C₀₋₄ alkyl)thiomorpholinyl, —O—(C₀₋₄alkyl)imidazolyl, —O—(C₀₋₄ alkyl)thienyl, —O—(C₀₋₄alkyl)tetrahydropyranyl, —O—(C₀₋₄ alkyl)tetrahydrofuranyl, —O—(C₀₋₄alkyl)pyrrolidinyl, —O—(C₀₋₄ alkyl)piperidinyl, or —O—(C₀₋₄alkyl)piperazinyl group;

R₅ and R₆ are independently —H, —OH, or an unsubstituted C₁₋₆ alkoxygroup; or R₄ and R₅ together are a methylenedioxy group, or R₅ and R₆together are a methylenedioxy group; and

R₈ is —H, —OH, —COOH, or an unsubstituted alkyl or —(CH₂)₁₋₆-phenylgroup.

In another embodiment, the invention provides a second group ofcompounds of Formula III,

stereoisomers thereof, tautomers thereof, solvates thereof, andpharmaceutically acceptable salts thereof; wherein

R₁ and R₂ are independently —H, —(CH₂)₀₋₆COOR′, —C(O)R″, or asubstituted or unsubstituted alkyl, cycloalkyl, cycloalkylalkyl,alkenyl, aryl, aralkyl, heteroaryl, heteroarylalkyl, heterocyclyl, orheterocyclylalkyl group; or R₁ and R₂ together are a methylene group;

R₃ and R₈ are independently —H, —OH, —Cl, —Br, —F, —I, —CN, —NH₂,—C(O)NH₂, —COOH, or a substituted or unsubstituted alkyl, alkenyl,alkoxy or aralkyl group;

R₃′ is —H, or R₃ and R₃′ together are an oxo group;

R₄ is —H, halogen, —OR′, —OSO₂R″, —OC(O)R″, —OC(O)OR″, —OC(O)NR′R″,—O-alkylene-NR′R′, —O-alkylene-OSO₂R″, —O-alkylene-S(O)₀₋₂R″,—O-alkylene-NR′SO₂R″, —O-alkylene-N(R′)C(O)R′, or a substituted orunsubstituted alkyl group;

R₅ and R₆ are independently —H, halogen, —OH, or a substituted orunsubstituted alkoxy group; or R₄ and R₅ together are a methylenedioxygroup, or R₅ and R₆ together are a methylenedioxy group;

R₇ is —H, halogen, —OH, or a substituted or unsubstituted alkyl oralkoxy group;

each R′ is independently a hydrogen, or a substituted or unsubstitutedalkyl, alkenyl, cycloalkyl, cycloalkylalkyl, aryl, aralkyl, heteroaryl,heteroarylalkyl, heterocyclyl, or heterocyclylalkyl group;

each R″ is independently a substituted or unsubstituted alkyl, alkenyl,cycloalkyl, cycloalkylalkyl, aryl, aralkyl, heteroaryl, heteroarylalkyl,heterocyclyl, or heterocyclylalkyl group;

with the proviso that when R₄ is —H, —OH or a C₁₋₄ alkoxy group, then R₅is not —H, —OH or a C₁₋₄ alkoxy group; and when R₁ and R₂ are both —CH₃or when R₁ and R₂ together are a methylene group, then R₅ is not OH or aC₁₋₂ alkoxy group, and R₄ and R₅ together are not a methylenedioxygroup; and when R₄ is OC(O)R″, then R₅ is not OC(O)R″ or methoxy.

In some embodiments of the first and second groups of compounds ofFormula III (collectively, “compounds of Formula III) and the compoundsof Formula IV, R₁ and R₂ are independently —H, —(CH₂)₀₋₂COOR′,—C(O)(CH₂)₀₋₂R″, or a unsubstituted C₁₋₆ alkyl group; or R₁ and R₂together are a methylene group. In other embodiments, R₁ and R₂ togetherare a methylene group.

In some embodiments of the compounds of Formula III and the compounds ofFormula IV, R₃ and R₃′ are each —H, or R₃ and R₃′ together are an oxogroup.

In some embodiments of compounds of Formula III and the compounds ofFormula IV, R₄ is —H, —OR′, —OSO₂R″, —OC(O)OR″, —OC(O)NR′R″,—O-alkylene-OSO₂R″, or —O-alkylene-NR′R′. In other embodiments, R₄ is—H, —OH, or a substituted or unsubstituted C₁₋₆ alkoxy, C₇₋₁₄ aralkoxy,—OC(O)—(C₁₋₆ alkyl), —OC(O)-(aryl), —OC(O)O-(aryl), —OC(O)—NH-(aryl),—O—(C₂₋₆ alkylene)-NH—(C₂₋₆ alkyl), —O—(C₂₋₆alkylene)-NH-(tetrahydropyran), —O—(C₂₋₆ alkylene)-NH-(thiomorpholinedioxide), —O—(C₂₋₆ alkylene)-NH-(piperidinyl), —O—(C₂₋₆alkylene)-NH-(piperazinyl), —O—(C₂₋₆ alkylene)-NH-(morpholinyl),—O—(C₂₋₆ alkylene)-NH-(aralkyl), —O—(C₂₋₆ alkylene)-NH-(cyclopropyl),—OSO₂—(C₃₋₆ cycloalkyl), —OSO₂-(aryl), —O—(C₂₋₆ alkylene)-OSO₂-(aryl),—OSO₂-(aralkyl), —O—(C₂₋₆ alkylene)-OSO₂-(heteroaryl), —OSO₂—(C₁₋₆alkyl), —OSO₂-(pyridyl), —OSO₂-(thiazolyl), —O—(C₂₋₆alkylene)-NHSO₂-(aryl), —O—(C₂₋₆ alkylene)-NHSO₂-(heteroaryl), —O—(C₂₋₆alkylene)-NHC(O)-(aryl), —O—(C₂₋₆ alkylene)-NHC(O)-(heteroaryl),—O—(C₀₋₄ alkyl)pyridyl, —O—(C₀₋₄ alkyl)pyrimidinyl, —O—(C₀₋₄alkyl)morpholinyl, —O—(C₀₋₄ alkyl)thiomorpholinyl, —O—(C₀₋₄alkyl)imidazolyl, —O—(C₀₋₄ alkyl)thienyl, —O—(C₀₋₄alkyl)tetrahydropyranyl, —O—(C₀₋₄ alkyl)tetrahydrofuranyl, —O—(C₀₋₄alkyl)pyrrolidinyl, —O—(C₀₋₄ alkyl)piperidinyl, or —O—(C₀₋₄alkyl)piperazinyl group.

In other embodiments of compounds of Formula III and compounds ofFormula IV, R₅ is OH or unsubstituted alkoxy and R₆ is H.

In some embodiments of compounds of Formula III and compounds of FormulaIV, R₈ is —H, —OH, —COOH, or an unsubstituted alkyl or —(CH₂)₁₋₆-phenylgroup.

In certain embodiments of compounds of Formula III and compounds ofFormula IV,

R₁ and R₂ are independently —H, —(CH₂)₀₋₂COOR′, —C(O)(CH₂)₀₋₂R″, or aunsubstituted C₁₋₆ alkyl group; or R₁ and R₂ together are a methylenegroup;

R₃ and R₃′ are each —H, or R₃ and R₃′ together are an oxo group;

R₄ is —H, —OH, or a substituted or unsubstituted C₁₋₆ alkoxy, C₇₋₁₄aralkoxy, —OC(O)—(C₁₋₆ alkyl), —OC(O)-(aryl), —OC(O)O-(aryl),—OC(O)—NH-(aryl), —O—(C₂₋₆ alkylene)-NH—(C₂₋₆ alkyl), —O—(C₂₋₆alkylene)-NH-(tetrahydropyran), —O—(C₂₋₆ alkylene)-NH-(thiomorpholinedioxide), —O—(C₂₋₆ alkylene)-NH-(piperidinyl), —O—(C₂₋₆alkylene)-NH-(piperazinyl), —O—(C₂₋₆ alkylene)-NH-(morpholinyl),—O—(C₂₋₆ alkylene)-NH-(aralkyl), —O—(C₂₋₆ alkylene)-NH-(cyclopropyl),—OSO₂—(C₃₋₆ cycloalkyl), —OSO₂-(aryl), —O—(C₂₋₆ alkylene)-OSO₂-(aryl),—OSO₂-(aralkyl), —O—(C₂₋₆ alkylene)-OSO₂-(heteroaryl), —OSO₂—(C₁₋₆alkyl), —OSO₂-(pyridyl), —OSO₂-(thiazolyl), —O—(C₂₋₆alkylene)-NHSO₂-(aryl), —O—(C₂₋₆ alkylene)-NHSO₂-(heteroaryl), —O—(C₂₋₆alkylene)-NHC(O)-(aryl), —O—(C₂₋₆ alkylene)-NHC(O)-(heteroaryl),—O—(C₀₋₄ alkyl)pyridyl, —O—(C₀₋₄ alkyl)pyrimidinyl, —O—(C₀₋₄alkyl)morpholinyl, —O—(C₀₋₄ alkyl)thiomorpholinyl, —O—(C₀₋₄alkyl)imidazolyl, —O—(C₀₋₄ alkyl)thienyl, —O—(C₀₋₄alkyl)tetrahydropyranyl, —O—(C₀₋₄ alkyl)tetrahydrofuranyl, —O—(C₀₋₄alkyl)pyrrolidinyl, —O—(C₀₋₄ alkyl)piperidinyl, or —O—(C₀₋₄alkyl)piperazinyl group;

R₅ and R₆ are independently —H, —OH, or an unsubstituted C₁₋₆ alkoxygroup; or R₄ and R₅ together are a methylenedioxy group, or R₅ and R₆together are a methylenedioxy group; and

R₈ is —H, —OH, —COOH, or an unsubstituted alkyl or —(CH₂)₁₋₆-phenylgroup.

In other embodiments of the compounds of Formula III and compounds ofFormula IV,

R₁ and R₂ are independently —H, —CH₃, —CH₂COOH, —CH₂C(O)OCH₂CH₃, allyl,or R₁ and R₂ together are a methylene group;

R₃ and R₃′ are each —H, or R₃ and R₃′ together are an oxo group;

R₄ is —H, —OH, OCH₃, —OCH₂CH₃, —O(CH₂)₂OH, —OCH₂COOH, —OCH₂COOCH₂CH₃,—O(CH₂)₂COOH, —O(CH₂)₂CH₂Br, —O-acetyl, —O-benzoyl,—O—(CH₂)₂—NH—(CH₂)₂—N(CH₃)₂, —O—(CH₂)₂—NH—(CH₂)₂—OCH₃,—O—(CH₂)₂—NH—(CH₂)₂—SCH₃, —O—(CH₂)₂—NH-morpholinyl,—O—(CH₂)₂—NH—(CH₂)₃—N(CH₃)₂, —O—(CH₂)₂—NH-benzyl,—O—(CH₂)₂—NH—(CH₂)₃-(thiomorpholine dioxide),—O—(CH₂)₂—NH—(CH₂)₃-morpholinyl, —O—(CH₂)₂—NH—(CH₂)₃-tetrahydropyranyl,—O-pyridyl optionally substituted with one or two substituents selectedfrom the group consisting of C₁₋₄ alkyl, —NO₂, and NH₂,—O—(CH₂)₂—S-phenyl, —OSO₂-naphthyl optionally substituted with di(C₁₋₄alkyl), —OSO₂—CF₃, —OSO₂-thiaolyl optionally substituted with acetamido,—O—(CH₂)₀₋₂SO₂-phenyl wherein the phenyl group is optionally substitutedwith one or two substituents selected from the group consisting ofmethyl, methoxy, fluoro, chloro, trifluoromethyl, and nitro,—OSO₂-cyclopentyl, —OSO₂-thienyl, —OSO₂-benzyl, —(CH₂)₂-cyclopropyl,—(CH₂)₂-morpholinyl, —(CH₂)₂-imidazolyl, —(CH₂)₂-pyrrolidinyl, or—(CH₂)₂-piperazinyl group, wherein the piperazinyl group is optionallysubstituted with methyl, isopropyl, or methoxyethyl;

R₅ and R₆ are independently —H, —OH, or —OCH₃; and

R₈ is —H, methyl, ethyl, —COOH, or benzyl.

While compounds of Formula III with either stereochemical configurationat position 14 exhibit lipid-lowering activity, the R-(+) stereochemicalconfiguration is generally preferred. Thus, compounds of Formula III canbe racemic at position 14 or can be a mixture of enantiomers having from1% to 99% enantiomeric excess (e.e.) with respect to the to R-(+)stereochemical configuration. For example, the compound of Formula IIImay have at least 1%, at least 5%, at least 10%, at least 20%, at least30%, at least 40%, at least 50%, at least 60%, at least 70%, at least80%, at least 90%, at least 95%, at least 96%, at least 97%, at least98%, or at least 99% e.e. Production and/or separation of either opticalisomer of compounds of Formula III is within the skill in the art inview of the guidance provided herein.

Likewise, certain compounds of Formula IV having the R-(+)stereochemical configuration at position 1 may exhibit improvedlipid-lowering activity compared to the opposite configuration at thisposition. In certain embodiments, the compound of Formula IV is anequimolar mixture of stereoisomers at position 1. As the compound ofFormula IV also has a stereocenter at position 9, two diastereomershaving the R-(+) stereochemical configuration at position 1 arepossible. In some embodiments, the compound of Formula IV has the(1R,9S) configuration. In other embodiments, the compound of Formula IVcan be a mixture of diastereomers having from 1% to 99% diastereomericexcess (d.e.) with respect to the to R-(+) stereochemical configurationat position 1. For example, the compound of Formula IV may have at least1%, at least 5%, at least 10%, at least 20%, at least 30%, at least 40%,at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, atleast 95%, at least 96%, at least 97%, at least 98%, or at least 99%d.e. with respect to position 1.

Compounds of Formulas I, II, III, and IV may be isolated from plants orprepared using the synthetic schemes described herein. Many suchcompounds may be made starting from natural products such as berberine.For example, Scheme 1 shows that berberine may be heated (e.g., 150-250°C.), preferably in a dry oven under reduced pressure, to selectivelyremove the position 19 (berberine numbering) methyl group and provideberberrubine.

The resulting hydroxyl group may be alkylated to provide product A. Thealkylation may be carried out with a wide variety of alkylating agentsR′X to provide various —OR′ wherein R′ is other than H such as alkyl,alkenyl, aralkyl, cycloalkyl, cycloalkylalkyl, heteroarylalkyl, andheterocyclylalkyl. X may be halides such as Cl, Br, or I, or X may beother leaving groups such as mesylate, trifluoromethanesulfunate,p-toluenesulfonate and the like. The alkylation may be carried out in asuitable solvent such as DMF, dichloromethane, chloroform or acetone bystirring or refluxing at a suitable temperature (e.g., ambient or withheating) until the desired product is formed. Optionally, a base is usedin the alkylation such as inorganic base (alkali metal carbonates) or anorganic base (pyridine, triethylamine).

In the third step, compound A may be reduced using any suitable reducingagent to give tetrahydroberberine compound B. Typically, borohydridesmay be used as the reducing agent, such as sodium borohydride, sodiumcyanoborohydride, or sodium triacetoxyborohydride. The reaction may becarried out in any suitable solvent or mixture of solvents (e.g.,alcohols such as methanol, ethanol, aqueous solutions thereof, andsolutions of AcOH) at a suitable temperature. It is within the skill inthe art to select a suitable temperature and reaction time for thereduction. Alternatively, the reduction may be carried out prior to thealkylation reaction (scheme not shown) so long as alkylation of the ringnitrogen is avoided.

In a similar fashion, as shown in Scheme 2, berberrubine may bealkylated with a terminal dihaloalkane X—(CH₂)_(n)—X, (n=1-10) to allowfor subsequent functionalization with an amine such as HNR′R″. Thealkylation reaction to give compound C may be carried out in a suitablesolvent such as DMF, dichloromethane, chloroform or acetone by stirringor refluxing at a suitable temperature until the reaction is complete.Amination of compound C with HNR′R″ (where R′ and R″ are as definedherein) to give compound D is optionally carried out in the presence ofan inorganic base (alkali metal carbonates) or an organic base(pyridine, triethylamine) in a suitable solvent (e.g., DMF,dichloromethane or chloroform), and at a suitable temperature. CompoundD is then reduced as described above to give compound E.

Similarly, as shown in Scheme 3, compound C in a suitable solvent may bereacted with various thiols (HSR″) to give compound F. As described forthe amination of C above, the reaction is optionally carried out in thepresence of an inorganic or organic base. Reduction as described hereinprovides the tetrahydroberberine derivative, compound G. The sulfone Hmay be prepared by exposing compound G to a mild oxidant such asperoxybenzoic acids (e.g., meta-chloroperoxybenzoic acid).

Scheme 4 shows a method for preparing acylated derivatives ofberberrubine. Compound 1 may be reduced as described herein to give atetrahydroberberine compound J. In the second step, compound J may beacylated with an acyl halide (e.g., R″C(O)X, where R″ is as definedherein and X is a halide such as Cl or Br), a haloformate (e.g.,R″OC(O)X), or an isocyanates (e.g., NC(O)R″) to provide, respectively,the corresponding amide, urethane or carbonate. The acylation istypically carried out in a suitable solvent in the presence of aninorganic base (alkali metal carbonates) or an organic base (pyridine,triethylamine). Upon completion of the reaction, the reaction mixture iscooled, and the product is optionally subjected to further separationand purification steps to give the target compound K. Likewise, R₄sulfonyl groups may be installed by reaction of compound J with asulfonyl halide, R″SO₂X, in the presence of an inorganic or organic basein a suitable solvent.

In another example of R₄ substituents, as shown in Scheme 5,berberrubine may be alkylated with a haloester (e.g., X(CH₂)nCOOR, whereX is a halo or other leaving group and R is a substituted orunsubstituted alkyl or aralkyl group) in a suitable solvent such asacetone, methanol, ethanol or mixtures thereof to give compound M. Thelatter compound may be reduced as described herein to give thetetrahydroberberine derivative N. The ester group may then be removed bystandard methods known in the art such alkaline or acid hydrolysis or,in the case of suitable aralkyl esters, by hydrogenolysis with asuitable catalyst (e.g. Pd/C, Pt/C, etc.). The compound O amide may beformed from the resulting acid by standard techniques such as reactingHNR′R″ in the presence of amide coupling reagents such as carbodiimides(e.g., DCC, EDC) in the presence of additives (HOBt, HOAt, DMAP), BOP,or by the formation of the corresponding acyl halide or mixed anhydride.

Compounds of Formulas I and III having various substituents at R₂ may beprepared by procedures analogous to those in schemes 1-4. Thus, forexample, as shown in Scheme 6, corypalmine may be alkylated with R₂X,wherein R₂ is as defined herein and X can be a halo, sulfonyl or otherleaving group under conditions described above. Similarly, the freehydroxyl in corypalmine may be acylated with an acyl halide in thepresence of a base or a carboxylic acid in presence of, for example, acoupling agent such as EDC.HCl/DMAP to give the target compound L.

R₈ substituents may be installed at the 13-position of compounds ofFormulas I and III as shown in Scheme 7. For example, an aqueoussolution of berberine chloride may be reacted with acetone in presenceof a suitable base such as alkali metal hydroxide to give the compoundQ. The protected compound Q can subsequently be reacted with R₇X,wherein R₇ is as described herein and X is a halide, sulfonyl group orother leaving group. The reaction is conducted in a suitable solvent ata suitable temperature optionally in presence of an alkali metal halidesuch as potassium iodide to give compound R. Compound R is hydrogenatedas described herein or with hydrogen using a suitable catalyst such asPt/C to give the tetrahydroberberine compound S.

Compounds of Formulas I and III may be made by total synthesis as shownin Scheme 8. The phenylacetic acid P may be coupled to thephenethylamine Q using standard techniques for the formation of amidebonds such as the use of coupling reagents (e.g., EDC/HOBt, carbonyldiimidazole, etc.), via formation of the acyl halide or mixed anhydrideof P. The resulting N-acyl β-arylethyl amine compound R may be subjectedto a Bischler-Napieralski reaction in a suitable solvent such asbenzene, toluene or xylene and in presence of a dehydrating agent suchas POCl₃ to give the corresponding dihydroisoquinoline compound S. Thelatter compound may be reduced by any suitable method such as withsodium borohydride, sodium cyanoborohydride or the like to give compoundT. Ring closure of compound T may be effected by reacting it withformaldehyde in a suitable solvent such as acetic acid to give thecompound U, which is a compound of Formula I and III.

Alternatively, Scheme 9 shows another general synthetic route tocompounds of Formulas I and III. Phenyl acetic acid derivative V may beexposed consecutively to phenylboronic acid, followed byparaformaldehyde. Both stages of the reaction are typically heated, andthe reaction with paraformaldehyde may be carried out under pressure in,e.g., a stainless steel bomb. Suitable solvents for this reactioninclude aromatic solvents such as toluene. The resulting boronate ishydrolyzed with water to give compound W. The latter compound may bealkylated with a wide variety of electrophiles, R′X, as described herein(e.g., for A in Scheme 1). Subsequently the amide may be formed with aphenethylamine compound as shown to give compound X. Ring closure usingPOCl₃ in a suitable solvent, such as toluene, followed by reduction asdescribed herein, gives compound Y, an exemplary compound of formulas Ior III.

Compounds of Formulas II and IV may be prepared according to Scheme 10.Thus, the starting phenethylamine with R₁ and R₂ already in place may bemade using standard techniques in the art. R₈ may be installed on thephenethylamine by reductive amination with an appropriate aldehyde(commercially available or prepared from the corresponding alcoholaccording to standard oxidation protocols). The startingcarboxytetrahydroisobenzofuran may also be readily prepared usingstandard techniques. The phenethylamine and thearboxytetrahydroisobenzofuran may be coupled using amide couplingreagents or other standard techniques. Thus, for example, coupling maybe effected in the presence of EDC/HOBt or carbonyl diimidazole amongother coupling reagents.

Another general synthetic route to compounds of Formulas II and IV isshown in Scheme 11. Compound A can be prepared throughBischler-Napieralsky cyclization of the corresponding phenethylamine,and can subsequently be reacted with lactone in the presence of a strongbase, such as LDA, to afford the precursor B. The latter compound may beconverted to various compounds of Formulas II and IV by reduction of thelactone carbonyl or reaction with, e.g., Grignard reagents to install R₃and R₃′.

Still other methods of preparing compounds of Formulas II and IV may beadapted from literature procedures as outlined in Scheme 12 anddescribed in the following references: Jerome L. Moniot and MauriceShamma, “Conversion of berberine into phthalideisoquinolines” J. Org.Chem., 1979, 44 (24), 4337-4342; Jerome L. Moniot, David M. Hindenlang,and Maurice Shamma, “Chemistry of 8,13-dioxoberbines” J. Org. Chem.,1979, 44(24), 4343-4346; Tatsuya Shono, Hiroshi Hamaguchi, Manji Sasaki,Shumei Fujita, and Kimihiko Nagami, “Novel zinc-promoted alkylation ofiminium salts. New synthesis of benzylisoquinoline,phthalidylisoquinoline, and protoberberine alkaloids and relatedcompounds” J. Org. Chem., 1983, 48(10), 1621-1628; R H Prager, J MTippett and A D Ward, “Central nervous system active compounds. VIII.New syntheses of phthalide isoquinolines” Aust. J. Chem., 1981, 34(5),1085-1093; SI Clarke, B Kasum, R H Prager and A D Ward, “Central nervoussystem active compounds. XIII. The use of aminomethylene phthalides inthe synthesis of phthalideisoquinoline alkaloids” Aust. J. Chem., 1983,36(12), 2493-2498; Nurani S. Narasimhan, Ravindra R. Joshi and (Mrs)Radhika S. Kusurkar, “An efficient synthesis of phthalideisoquinolinealkaloids” J. Chem. Soc., Chem. Comm., 1985, 3, 177-178; YoshikazuKondo, Jiro Imai and Shigeo Nozoe, Reaction of protoberberine-typealkaloids. Part 13. Biogenetic conversion of protoberberine alkaloidsinto phthalideisoquinoline alkaloids” J. Chem. Soc., Perkin. Trans. 1,1980, 919-926; Tetsuji Kametani, Toshio Honda, Hitoshi Inoue, andKeiichiro Fukumoto, “A One Step Synthesis of the PhthalideisoquinolineAlkaloid Cordrastine” Heterocycl. 1975, 3(12), 1091-1098; K. W. Bentleyand A. W. Murray, “Ketolaudanosine” J. Chem. Soc., 1963, 2487-2491.

For example, Scheme 13 shows how compound B may be prepared as reportedin Aust. J. Chem. 1983, 36(12), 2493. Compound D, prepared from3H-isobenzofuran-1-one (A) in two steps, can react with variousalkylating and acylating agents in the presence of base to afford theintermediate B.

In another aspect, the invention provides plant extracts containingisoquinolinyl alkaloids having lipid-lowering activity. For example,extracts from a number of species of Corydalis are suitable for use inthe methods described herein, including: ambigua, bulbosa, cava,chaerophylla, pallida, solida, thalictrifolia, tuberosa, andturtschaminowii Besser. In addition, extracts from species known inChina as Corydalis yan hu suo and Corydalis Xiar R1 Wu may be used.Alkaloids that may be isolated from Corydalis for use in the presentmethods include (+)-corypalmine, 14R,13S-(+)-corydaline,14R-(+)tetrahydropalmitine, (+)-corlumidine, (+)-bicuculline, and(+)-egenine. Such alkaloids may be isolated in partially orsubstantially pure form according to methods well known in the art.Further lipid lowering agents found among the Corydalis alkaloids andsuitable for use include coptisine, isocorypalmine, (+)-corybulbine,(+)-thalictricavine, (+)-thalictrifoline, cavidine, and apocavidine.

Additional sources for the lipid lowering agents of the presentinvention are found in extracts of the following plants: Berberidaceaefamily, including Leontice leontopatalum (THP) and Mahonia aquifolium(corypalmine); Fumariaceae family, including F. vaillantii((+)-stylopine); Menispermaceae family, including Legnephora moorii(dehydrocorydalmine) and Stephania glabra (Roxb.) (corydalmine, THP);Papaveraceae family, including Chelidonium majus L. (corysamine,stylopine), Hunnemannia fumariaefolia Sweet (corysamine), C. meifolia.(apocavidine, (+)-cavidine), and D. leptopodum (corysamine);Rannunculaceae family, including Coptis groenlandica (coptisine);Annonaceae family, including Guatteria discolor (corypalmine) andPachypodanthium staudtii (corypalmine); Menispermaceae family, includingChasmanthera dependens (THP) and Fibraurea chloroleuca (THP); G.vitellinum (THP).

In another aspect, the instant invention provides pharmaceuticalcompositions and medicaments comprising any of the compounds or extractsdisclosed herein (e.g., compounds of Formulas I, II, III, or IV) and apharmaceutically acceptable carrier or one or more excipients orfillers. In some embodiments, there are provided pharmaceuticalcompositions for treating a condition selected from the group consistingof hyperlipidemia, hypercholesterolemia, hypertriglyceridemia, hepaticsteatosis, and metabolic syndrome. Such compositions include alipid-lowering effective amount of compound of Formula III or FormulaIV, as described herein. In one embodiment, the pharmaceuticalcomposition is packaged in unit dosage form. The unit dosage form iseffective in lowering lipid levels (e.g., at least one of totalcholesterol, LDL-cholesterol, triglyceride, and unesterified long chainfatty acids) in the bloodstream and/or in the liver when administered toa subject in need thereof.

The pharmaceutical compositions may be prepared by mixing one or morecompounds of the invention, pharmaceutically acceptable salts thereof,stereoisomers thereof, tautomers thereof, or solvates thereof, withpharmaceutically acceptable carriers, excipients, binders, diluents orthe like to prevent and treat disorders associated with the effects ofincreased plasma and/or hepatic lipid levels. The compounds andcompositions described herein may be used to prepare formulations andmedicaments that prevent or treat a variety of disorders associated withincreased plasma and/or hepatic lipid levels, e.g., hyperlipidemia,hypercholesterolemia, hepatic steatosis, and metabolic syndrome. Suchcompositions can be in the form of, for example, granules, powders,tablets, capsules, syrup, suppositories, injections, emulsions, elixirs,suspensions or solutions. The instant compositions can be formulated forvarious routes of administration, for example, by oral, parenteral,topical, rectal, nasal, vaginal administration, or via implantedreservoir. Parenteral or systemic administration includes, but is notlimited to, subcutaneous, intravenous, intraperitoneal, andintramuscular, injections. The following dosage forms are given by wayof example and should not be construed as limiting the instantinvention.

For oral, buccal, and sublingual administration, powders, suspensions,granules, tablets, pills, capsules, gelcaps, and caplets are acceptableas solid dosage forms. These can be prepared, for example, by mixing oneor more compounds of the instant invention, or pharmaceuticallyacceptable salts or tautomers thereof, with at least one additive suchas a starch or other additive. Suitable additives are sucrose, lactose,cellulose sugar, mannitol, maltitol, dextran, starch, agar, alginates,chitins, chitosans, pectins, tragacanth gum, gum arabic, gelatins,collagens, casein, albumin, synthetic or semi-synthetic polymers orglycerides. Optionally, oral dosage forms can contain other ingredientsto aid in administration, such as an inactive diluent, or lubricantssuch as magnesium stearate, or preservatives such as paraben or sorbicacid, or anti-oxidants such as ascorbic acid, tocopherol or cysteine, adisintegrating agent, binders, thickeners, buffers, sweeteners,flavoring agents or perfuming agents. Tablets and pills may be furthertreated with suitable coating materials known in the art.

Liquid dosage forms for oral administration may be in the form ofpharmaceutically acceptable emulsions, syrups, elixirs, suspensions, andsolutions, which may contain an inactive diluent, such as water.Pharmaceutical formulations and medicaments may be prepared as liquidsuspensions or solutions using a sterile liquid, such as, but notlimited to, an oil, water, an alcohol, and combinations of these.Pharmaceutically suitable surfactants, suspending agents, emulsifyingagents, may be added for oral or parenteral administration.

As noted above, suspensions may include oils. Such oils include, but arenot limited to, peanut oil, sesame oil, cottonseed oil, corn oil andolive oil. Suspension preparation may also contain esters of fatty acidssuch as ethyl oleate, isopropyl myristate, fatty acid glycerides andacetylated fatty acid glycerides. Suspension formulations may includealcohols, such as, but not limited to, ethanol, isopropyl alcohol,hexadecyl alcohol, glycerol and propylene glycol. Ethers, such as butnot limited to, poly(ethyleneglycol), petroleum hydrocarbons such asmineral oil and petrolatum; and water may also be used in suspensionformulations.

Injectable dosage forms generally include aqueous suspensions or oilsuspensions which may be prepared using a suitable dispersant or wettingagent and a suspending agent. Injectable forms may be in solution phaseor in the form of a suspension, which is prepared with a solvent ordiluent. Acceptable solvents or vehicles include sterilized water,Ringer's solution, or an isotonic aqueous saline solution.Alternatively, sterile oils may be employed as solvents or suspendingagents. Typically, the oil or fatty acid is non-volatile, includingnatural or synthetic oils, fatty acids, mono-, di- or tri-glycerides.

For injection, the pharmaceutical formulation and/or medicament may be apowder suitable for reconstitution with an appropriate solution asdescribed above. Examples of these include, but are not limited to,freeze dried, rotary dried or spray dried powders, amorphous powders,granules, precipitates, or particulates. For injection, the formulationsmay optionally contain stabilizers, pH modifiers, surfactants,bioavailability modifiers and combinations of these.

Compounds of the invention may be administered to the lungs byinhalation through the nose or mouth. Suitable pharmaceuticalformulations for inhalation include solutions, sprays, dry powders, oraerosols containing any appropriate solvents and optionally othercompounds such as, but not limited to, stabilizers, antimicrobialagents, antioxidants, pH modifiers, surfactants, bioavailabilitymodifiers and combinations of these. The carriers and stabilizers varywith the requirements of the particular compound, but typically includenonionic surfactants (Tweens, Pluronics, or polyethylene glycol),innocuous proteins like serum albumin, sorbitan esters, oleic acid,lecithin, amino acids such as glycine, buffers, salts, sugars or sugaralcohols. Aqueous and nonaqueous (e.g., in a fluorocarbon propellant)aerosols are typically used for delivery of inventive compounds byinhalation.

Dosage forms for the topical (including buccal and sublingual) ortransdermal administration of compounds of the invention includepowders, sprays, ointments, pastes, creams, lotions, gels, solutions,and patches. The active component may be mixed under sterile conditionswith a pharmaceutically-acceptable carrier or excipient, and with anypreservatives, or buffers, which may be required. Powders and sprays canbe prepared, for example, with excipients such as lactose, talc, silicicacid, aluminum hydroxide, calcium silicates and polyamide powder, ormixtures of these substances. The ointments, pastes, creams and gels mayalso contain excipients such as animal and vegetable fats, oils, waxes,paraffins, starch, tragacanth, cellulose derivatives, polyethyleneglycols, silicones, bentonites, silicic acid, talc and zinc oxide, ormixtures thereof. Absorption enhancers can also be used to increase theflux of the inventive compound across the skin. The rate of such fluxcan be controlled by either providing a rate controlling membrane (e.g.,as part of a transdermal patch) or dispersing the compound in a polymermatrix or gel.

Besides those representative dosage forms described above,pharmaceutically acceptable excipients and carriers are generally knownto those skilled in the art and are thus included in the instantinvention. Such excipients and carriers are described, for example, in“Remingtons Pharmaceutical Sciences” Mack Pub. Co., New Jersey (1991),which is incorporated herein by reference.

The formulations of the invention may be designed to be short-acting,fast-releasing, long-acting, and sustained-releasing as described below.Thus, the pharmaceutical formulations may also be formulated forcontrolled release or for slow release.

The instant compositions may also comprise, for example, micelles orliposomes, or some other encapsulated form, or may be administered in anextended release form to provide a prolonged storage and/or deliveryeffect. Therefore, the pharmaceutical formulations and medicaments maybe compressed into pellets or cylinders and implanted intramuscularly orsubcutaneously as depot injections or as implants such as stents. Suchimplants may employ known inert materials such as silicones andbiodegradable polymers.

Specific dosages may be adjusted depending on conditions of disease, theage, body weight, general health conditions, sex, and diet of thesubject, dose intervals, administration routes, excretion rate, andcombinations of drugs. Any of the above dosage forms containingeffective amounts are well within the bounds of routine experimentationand therefore, well within the scope of the instant invention.

Those skilled in the art are readily able to determine an effectiveamount by simply administering a compound of the invention to a patientin increasing amounts until the elevated plasma or hepatic cholesterolor triglycerides or progression of the disease state is decreased orstopped. The progression of the disease state can be assessed using invivo imaging, as described, or by taking a tissue sample from a patientand observing the target of interest therein. The compounds of theinvention can be administered to a patient at dosage levels in the rangeof about 0.1 to about 1,000 mg per day. For a normal human adult havinga body weight of about 70 kg, a dosage in the range of about 0.01 toabout 100 mg per kg of body weight per day is sufficient. The specificdosage used, however, can vary or may be adjusted as consideredappropriate by those of ordinary skill in the art. For example, thedosage can depend on a number of factors including the requirements ofthe patient, the severity of the condition being treated and thepharmacological activity of the compound being used. The determinationof optimum dosages for a particular patient is well known to thoseskilled in the art.

Various assays and model systems can be readily employed to determinethe therapeutic effectiveness of antihyperlipidemia treatment accordingto the invention. For example, blood tests to measure total cholesterolas well as triglycerides, LDL and HDL levels are routinely given.Individuals with a total cholesterol level of greater than 200 mg/dL areconsidered borderline high risk for cardiovascular disease. Those with atotal cholesterol level greater than 239 mg/dL are considered to be athigh risk. An LDL level of less than 100 mg/dL is considered optimal.LDL levels between 130 to 159 mg/dL are borderline high risk. LDL levelsbetween 160 to 189 mg/dL are at high risk for cardiovascular disease andthose individuals with an LDL greater than 190 mg/dL are considered tobe at very high risk for cardiovascular disease. Triglyceride levels ofless than 150 mg/dL is considered normal. Levels between 150-199 mg/dLare borderline high and levels above 200 mg/dL are considered to put theindividual at high risk for cardiovascular disease. Lipid levels can bedetermined by standard blood lipid profile tests. Effective amounts ofthe compositions of the invention will lower elevated lipid levels by atleast 10%, 20%, 30%, 50% or greater reduction, up to a 75-90%, or 95% orgreater. Effective amounts will also move the lipid profile of anindividual towards the optimal category for each lipid, i.e., decreaseLDL levels from 190 mg/dL to within 130 to 159 mg/dL or even further tobelow 100 mg/dL. Effective amounts may further decrease LDL ortriglyceride levels by about 10 to about 70 mg/dL, by about 20 to about50 mg/dL, by about 20 to about 30 mg/dL, or by about 10 to about 20mg/dL.

A variety of hyperlipidemia classification systems are known to personsof skill in the art. One such classification system is the Fredericksonclassification, which is summarized in Table 1 below.

TABLE 1 Relative Elevated Elevated Plasma Plasma Frequency PhenotypeLipoproteins Lipid Levels TC TG (%)* I Chylomicrons TG N to ↑ ↑↑↑↑ <1IIa LDL TC ↑↑ N 10 IIb LDL & TC, TG ↑↑ ↑↑↑ 40 VLDL III IDL TC, TG ↑↑ ↑↑<1 IV VLDL TG, TC N to ↑ ↑↑ 45 V VLDL & TG, TC ↑to ↑↑ ↑↑↑↑ 5 chylomicronIDL, intermediate-density lipoprotein; LDL, low-density lipoprotein; N,normal; TC, total cholesterol; TG, triglyceride; VLDL, very-low-densitylipoprotein *Approximate % of patients in the United States withhyperlipidemia.

Individuals may also be evaluated using a hs-CRP (high-sensitivityC-reactive protein) blood test. Those with a hs-CRP result of less than1.0 mg/L are at low risk for cardiovascular disease. Individuals with ahs-CRP result between about 1.0 to 3.0 mg/L are at average risk forcardiovascular disease. Those with a hs-CRP result greater than 3.0 mg/Lare at high risk of cardiovascular disease. Effective amounts of thecompositions of the present invention will lower hs-CRP results below3.0 mg/L. Effective amounts of the compositions of the present inventioncan lower hs-CRP results by about 0.5 to about 3.0 mg/L, and further byabout 0.5 to about 2.0 mg/L.

Effectiveness of the compositions and methods of the invention may alsobe demonstrated by a decrease in the symptoms of cardiovascular disease,edema, diabetes insipidus, hypertension, myocardial ischemia, congestiveheart failure, arrhythmia, and hyperlipoproteinemia, the symptomsincluding shortness of breath, chest pain, leg pain, tiredness,confusion, vision changes, blood in urine, nosebleeds, irregularheartbeat, loss of balance or coordination, weakness, or vertigo.

For each of the indicated conditions described herein, test subjectswill exhibit a 10%, 20%, 30%, 50% or greater reduction, up to a 75-90%,or 95% or greater, reduction, in one or more symptom(s) caused by, orassociated with, hyperlipidemia, elevated cholesterol, elevatedtriglyceride, and/or a targeted cardiovascular disease or condition inthe subject, compared to placebo-treated or other suitable controlsubjects.

The compounds of the invention can also be administered to a patientalong with other conventional therapeutic agents that may be useful inthe treatment or prophylaxis of hyperlipidemic diseases. In one aspect,a method is provided for administering an effective amount of one ormore compounds of the invention to a patient suffering from or believedto be at risk of suffering from a disease characterized by elevatedplasma or hepatic cholesterol or triglycerides. Moreover, the inventionrelates to treating a hyperlipidemic disease by administering aneffective amount of one or more compounds to a patient in need thereof.The methods of the invention can also comprise administering, eithersequentially or in combination with one or more compounds of theinvention, a conventional therapeutic agent in an amount that canpotentially or synergistically be effective for the treatment orprophylaxis of a hyperlipidemic disease. Exemplary therapeutic agentsfor use in combination therapies with one or more compounds of theinvention include, but are not limited to, anti-inflammatory drugs,therapeutic antibodies and cholesterol lowering drugs such as, forexample, statins.

In one aspect, a compound of the invention is administered to a patientin an amount or dosage suitable for therapeutic use. Generally, a unitdosage comprising a compound of the invention will vary depending onpatient considerations. Such considerations include, for example, age,protocol, condition, sex, extent of disease, contraindications,concomitant therapies and the like. An exemplary unit dosage based onthese considerations can also be adjusted or modified by a physicianskilled in the art. For example, a unit dosage for a patient comprisinga compound of the invention can vary from 1×10⁻⁴ g/kg to 1 g/kg,preferably, 1×10⁻³ g/kg to 1.0 g/kg. Dosage of a compound of theinvention can also vary from 0.01 mg/kg to 100 mg/kg or, preferably,from 0.1 mg/kg to 10 mg/kg.

Useful adjunctive therapeutic agents in combinatorial formulations andcoordinate treatment methods include, for example, antihyperlipidemicagents; antidyslipidemic agents; antidiabetic agents, including, but notlimited to metformin, rosiglitazone, plasma HDL-raising agents,including, but not limited to, nicotinic acid, fibrates;antihypercholesterolemic agents, including, but not limited to,cholesterol-uptake inhibitors; cholesterol biosynthesis inhibitors,e.g., HMG-CoA reductase inhibitors (also referred to as statins, such aslovastatin, simvastatin, pravastatin, fluvastatin, rosuvastatin,pitavastatin, and atorvastatin); HMG-CoA synthase inhibitors; squaleneepoxidase inhibitors or squalene synthetase inhibitors (also known assqualene synthase inhibitors); microsomal triglyceride transfer protein(MTP) inhibitor; acyl-coenzyme A cholesterol acyltransferase (ACAT)inhibitors, including, but not limited to, melinamide; probucol;nicotinic acid and the salts thereof; niacinamide; cholesterolabsorption inhibitors, including, but not limited to, beta-sitosterol orezetimibe; bile acid sequestrant anion exchange resins, including, butnot limited to cholestyramine, colestipol, colesevelam ordialkylaminoalkyl derivatives of a cross-linked dextran; LDL receptorinducers; fibrates, including, but not limited to, clofibrate,bezafibrate, fenofibrate and gemfibrozil; vitamin B6 (pyridoxine) andthe pharmaceutically acceptable salts thereof, such as the HCl salt;vitamin B12 (cyanocobalamin); vitamin B3 (nicotinic acid andniacinamide); anti-oxidant vitamins, including, but not limited to,vitamin C and E and beta carotene; beta blockers; angiotensin IIreceptor (AT₁) antagonist; angiotensin-converting enzyme inhibitors,renin inhibitors; platelet aggregation inhibitors, including, but notlimited to, fibrinogen receptor antagonists, i.e., glycoprotein Ib/IIIafibrinogen receptor antagonists; hormones, including but not limited to,estrogen; insulin; ion exchange resins; omega-3 oils; benfluorex; ethylicosapentate; and amlodipine. Adjunctive therapies may also includeincrease in exercise, surgery, and changes in diet (e.g., to a lowcholesterol diet). Some herbal remedies may also be employed effectivelyin combinatorial formulations and coordinate therapies for treatinghyperlipidemia, for example curcumin, gugulipid, garlic, soy, solublefiber, fish oil, green tea, carnitine, chromium, coenzyme Q10, grapeseed extract, pantothine, red yeast rice, and royal jelly.

Berberine and related compounds also can be employed as secondtherapeutic agents together with the Corydalis lipid lowering agents ofthe invention. For example, berberine sulfate, berberine hydrochloride,berberine chloride, oxyberberine, dihydroberberine,8-cyanodihydroberberine, tetrahydroberberine N-oxide,tetrahydroberberine, 6-protoberberine, 9-ethoxycarbonyl berberine,9-N,N-dimethylcarbamoyl berberine and 12-bromo berberine, berberineazide, and berberine betaine can be used. Berberine compounds that areeffective in raising the expression level of LDLR are described in US2006/0223838, which is hereby incorporated by reference in its entirety.

Another class of compounds that can be used as second therapeutic agentstogether with the Corydalis lipid lowering agents of the invention isthe SCAP antagonists. These compounds bind to SREBP-cleavage activatingprotein and prevent its physical interaction with SREBP, resulting inactivation of the LDLR promoter and increased expression of LDLR.Suitable compounds are described in U.S. Pat. No. 6,673,555 (which ishereby incorporated by reference in its entirety).

In some embodiments a Corydalis lipid lowering agent is combined withone or more sterol 14-reductase inhibitors as second agents. Suchinhibitors will reduce the synthesis of cholesterol in the liver, andconsequently contribute to the reduction of total cholesterol andLDL-cholesterol. A series of suitable 14-reductase inhibitors based onCorydalis alkaloids is described in U.S. Pat. Nos. 6,255,317 and6,239,139, both of which are incorporated by reference in theirentirety. It is noteworthy that the Corydalis alkaloids which functionas 14-reductase inhibitors differ from the Corydalis lipid loweringagents of the present invention in having a double bond at the 13-14position. In some embodiments of the present invention, however, theadditional effect of inhibiting cholesterol synthesis may be undesired.In such cases, 14-reductase inhibitors, particularly those Corydalisalkaloids having a double bond at the 13-14 position, are specificallyexcluded from use with a Corydalis lipid lowering agent of the presentinvention.

A compound of the invention can bind to one or more targets of interestwith a dissociation constant (for example, an equilibrium dissociationconstant, K_(d)) from, for example, about 0.0001 to 10 μM (or from0.0001 to 7 μM, 0.0001 to 5 μM, 0.0001 to 1 μM, 0.001 to 5 μM, 0.01 to 5μM and/or 0.1 to 5 μM) as measured by any suitable techniques routine tothose of ordinary skill in the art. The invention contemplatesmeasurement of a dissociation constant (for example, K_(d) and K_(i)) orperforming competition, saturation and kinetics experiments byconventional techniques routine to one of ordinary skill in the art.Moreover, a compound of the invention can compete with a referencecompound for binding to and/or with targets of interest with adissociation constant of inhibition (for example, K_(i)) from, forexample, about 0.01 nM to >10,000 nM (or from 0.001 to 7,000 nM, 0.001to 5,000 nM, 0.001 to 1,000 nM, 0.01 to 5,000 nM, 0.01 to 2,000 nMand/or 0.1 to 5,000 nM).

A compound or probe of the invention can bind to one or more targets ofinterest with a dissociation constant (for example, an equilibriumdissociation constant, K_(d)) from, for example, about 0.0001 to 10 μMas measured by binding to a synthetic peptide or tissue associated witha target of interest. The invention contemplates measurement of adissociation constant (for example, K_(d) and K_(i)) or performingcompetition, saturation and kinetics experiments by conventionaltechniques routine to one of ordinary skill in the art. Moreover, acompound or probe of the invention can compete with a reference compoundfor binding to a target of interest with a dissociation constant ofinhibition (for example, K_(i)) from, for example, about 0.01 nMto >10,000 nM.

In one aspect, binding, interaction or association with can mean thecontact between a compound (or analogs, salts, pharmaceuticalcompositions, derivatives, metabolites, prodrugs or racemic, tautomersmixtures thereof) and a target of interest with a binding affinity of atleast 10⁻⁶ M, preferably, at least about 10⁻⁷ M, and more preferably10⁻⁸ M to 10⁻⁹ M, 10⁻¹⁰ M, 10⁻¹¹ M, or 10⁻¹² M. In one aspect, bindingaffinities include those with a dissociation constant or K_(d) lessthan, but not limited to, 5×10⁻⁶ M, 10⁻⁶ M, 5×10⁻⁷ M, 10⁻⁷ M, 5×10⁻⁸ M,10⁻⁸ M, 5×10⁻⁹ M, 10⁻⁹ M, 5×10⁻¹⁰ M, 10⁻¹⁰ M, 5×10⁻¹¹ M, 10⁻¹¹ M,5×10⁻¹² M, 10⁻¹² M, 5×10⁻¹³M, 10−13 M, 5×10⁻¹⁴ M, 10⁻¹⁴ M, 5×10⁻¹⁵ M,and 10⁻¹⁵ M.

A compound of the invention can also be modified, for example, by thecovalent attachment of an organic moiety or conjugate to improvepharmacokinetic properties, toxicity or bioavailability (e.g., increasedin vivo half-life). The conjugate can be a linear or branchedhydrophilic polymeric group, fatty acid group or fatty acid ester group.A polymeric group can comprise a molecular weight that can be adjustedby one of ordinary skill in the art to improve, for example,pharmacokinetic properties, toxicity or bioavailability. Exemplaryconjugates can include a polyalkane glycol (e.g., polyethylene glycol(PEG), polypropylene glycol (PPG)), carbohydrate polymer, amino acidpolymer or polyvinyl pyrolidone and a fatty acid or fatty acid estergroup, each of which can independently comprise from about eight toabout seventy carbon atoms. Conjugates for use with a compound of theinvention can also serve as linkers to, for example, any suitablesubstituents or groups, radiolabels (marker or tags), halogens,proteins, enzymes, polypeptides, other therapeutic agents (for example,a pharmaceutical or drug), nucleosides, dyes, oligonucleotides, lipids,phospholipids and/or liposomes. In one aspect, conjugates can includepolyethylene amine (PEI), polyglycine, hybrids of PEI and polyglycine,polyethylene glycol (PEG) or methoxypolyethylene glycol (mPEG). Aconjugate can also link a compound of the invention to, for example, alabel (fluorescent or luminescent) or marker (radionuclide, radioisotopeand/or isotope) to comprise a probe of the invention. Conjugates for usewith a compound of the invention can, in one aspect, improve in vivohalf-life. Other exemplary conjugates for use with a compound of theinvention as well as applications thereof and related techniques includethose generally described by U.S. Pat. No. 5,672,662, which is herebyincorporated by reference herein.

In another aspect, the invention provides methods of identifying atarget of interest including contacting the target of interest with adetectable or imaging effective quantity of a labeled compound of theinvention. A detectable or imaging effective quantity is a quantity of alabeled compound of the invention necessary to be detected by thedetection method chosen. For example, a detectable quantity can be anadministered amount sufficient to enable detection of binding of thelabeled compound to a target of interest including, but not limited to,one or more cellular proteins. Suitable labels are known by thoseskilled in the art and can include, for example, radioisotopes,radionuclides, isotopes, fluorescent groups, biotin (in conjunction withstreptavidin complexation), and chemoluminescent groups. Upon binding ofthe labeled compound to the target of interest, the target may beisolated, purified and further characterized such as by determining theamino acid sequence.

The terms “associated” and/or “binding” can mean a chemical or physicalinteraction, for example, between a compound of the invention and atarget of interest. Examples of associations or interactions includecovalent bonds, ionic bonds, hydrophilic-hydrophilic interactions,hydrophobic-hydrophobic interactions and complexes. Associated can alsorefer generally to “binding” or “affinity” as each can be used todescribe various chemical or physical interactions. Measuring binding oraffinity is also routine to those skilled in the art. For example,compounds of the invention can bind to or interact with a target ofinterest or precursors, portions, fragments and peptides thereof and/ortheir deposits.

The examples herein are provided to illustrate advantages of the presentinvention and to further assist a person of ordinary skill in the artwith preparing or using the compounds of the invention or salts,pharmaceutical compositions, derivatives, metabolites, prodrugs, racemicmixtures or tautomeric forms thereof. The examples herein are alsopresented in order to more fully illustrate the preferred aspects of theinvention. The examples should in no way be construed as limiting thescope of the invention, as defined by the appended claims. The examplescan include or incorporate any of the variations, aspects or aspects ofthe invention described above. The variations, aspects or aspectsdescribed above may also further each include or incorporate thevariations of any or all other variations, aspects or aspects of theinvention.

EXAMPLES Example 1 Isolation and Purification of 13S,14R-Corydaline

All chemicals were purchased from the Sigma-Aldrich Chemical Company(Milwaukee, Wis.). Solvents were purchased from VWR International(Brisbane, Calif.) and were all of HPLC purity standard or higher.Proton and ¹³C NMR spectra were performed on a 300 MHz Bruker AC-300plus NMR spectrometer with a TCPLink PC upgrade (INAC Computer, GmbH,Malsch, Germany). The NMR solvent was CDCl₃ unless otherwise specified.HPLC was performed using Waters 600 pumps and controller with a Waters996 photodiode array detector. Solvent A was 0.05% trifluoroacetic acidin water. Solvent B was 0.04% trifluoroacetic acid in acetonitrile. Thegradient was 0 to 100% B over 30 minutes, 2 mL/min. flow rate. Thecolumn was a C-18 reverse phase Vydac 254TP18 column of 25×0.46 cm.Flash chromatography was performed on a Teledyne Isco (Lincoln, Nebr.)CombiFlash Companion automated workstation. FT-IR spectra were obtainedon a Perkin-Elmer FT-1600 spectrophotometer, and melting points weredetermined on a Cole Palmer Kofler block melting point apparatus. X-raycrystallography for absolute configuration was performed at the Centerfor Chemical Characterization and Analysis, Texas A&M University(College Stations, Tex.).

It was determined by thin layer chromatography (1:1 hexane/ethyl acetateon normal phase silica plates then stained with iodine) that crudecorydaline purchased from Sigma-Aldrich Corporation was in fact, amixture of mainly corydaline (R_(f)=0.7) and a small amount of anunknown impurity (R_(f)=0.3) (see FIG. 5). Crude corydaline (250 mg) wasthen subjected to normal phase preparative flash chromatography. Thecrude material was loaded onto a 12 g Isco pre-packed silica column,eluted with 1:1 hexane/ethyl acetate, and 20 mL fractions werecollected. Fractions 14 to 16 were pooled and collected. Removal of theeluting solvent in vacuo, afforded an off-white powder (100 mg), whichwas recrystallized from ethyl acetate I hexane to give13S,14R-corydaline as off-white needles (m.p. 135 to 137° C.). Thismaterial was shown to be greater than 99% pure by reverse-phase HPLC(see FIG. 4). The proton NMR, ¹³C NMR, and mass spectra of this materialwere consistent with the structure of corydaline, and X-raycrystallography demonstrated that the absolute stereochemistry of thismaterial was in fact 13S,14R-corydaline (see Example 5).

Example 2 Isolation and Purification of 14R-Tetrahydropalmatine(14R-THP)

From the silica column of Example 1, the later eluting fractions 22 to26 containing the unknown impurity were pooled and collected. Thesolvent was removed in vacuo to afford 3 mg of a yellow powder. This wasrecrystallized from ethyl acetate/hexane to afford yellow crystals of14R-tetrahydropalmatine (2 mg). This material was shown to be greaterthan 95% pure by reverse-phase HPLC. The proton NMR spectrum of thismaterial showed that it was tetrahydropalmatine (THP), and X-raycrystallography demonstrated that the absolute stereochemistry was14R-THP (see Example 6).

Example 3 Synthesis of 14R-Tetrahydropalmatine (THP) from Berberine(BBR)

14R-THP was prepared from BBR in four steps (see scheme below) startingby treating BBR with boron trichloride in methylene chloride. Thisdeprotected only the methylene bridged catechol leaving the methoxygroups untouched. Methylation with methyl iodide and potassium carbonatein dry acetone then afforded the tetra-O-OMe compound that wassubsequently subjected to asymmetric hydrogenation with a suitableasymmetric hydrogenation catalyst to afford 14R-THP. The S-enantiomermay be similarly obtained. In addition, acid addition salts of 14R-THPmay be prepared by exposure to acid during the hydrogenation orafterwards as a separate step.

Exemplary catalysts that can be used for the synthesis are generallydescribed by: Bunlaksananusorn, T., Polbom, K., Knochel, P., “New P,Nligands for asymmetric Ir-catalyzed reactions,” Angew. Chemie, Intl. Ed.(2003), 42(33), 941-3943; Lu, S.-M., Han, X.-W., Zhou, Y.-G.,“Asymmetric hydrogenation of quinolines catalyzed by iridium with chiralferrocenyloxazoline derived N,P ligands,” Advanced Synthesis & Catalysis(2004), 346(8), 909-912; Lu, S.-M., Wang, Y.-Q., Han, X.-W., Zhou,Y.-G., “Asymmetric hydrogenation of quinolines and isoquinolinesactivated by chloroformates,” Angew. Chemie, Intl. Ed. (2006), 45(14),2260-2263; Wang, D.-W., Zeng, W.; Zhou, Y.-G., “Iridium-catalyzedasymmetric transfer hydrogenation of quinolines with Hantzsch esters,”Tetrahedron: Asymmetry (2007), 18(9), 1103-1107; Xu Lijin; Lam Kim Hung;Ji Jianxin; Wu Jing; Fan Qing-Hua; Lo Wai-Hung; Chan Albert S C“Air-stable Ir—(P-Phos) complex for highly enantioselectivehydrogenation of quinolines and their immobilization in poly(ethyleneglycol) dimethyl ether (DMPEG),” Chem. Comm. (Cambridge, England)(2005), (11), 1390-2; and Wang, Y., Weissensteiner, W., Spindler, F.,Arion, V. B., and Mereiter, K., “Synthesis and Use in AsymmetricHydrogenations of Solely Planar Chiral 1,2-Disubstituted and1,2,3-Trisubstituted Ferrocenyl Diphosphines: A Comparative Study,”Organometallics, 2007, each of which is incorporated herein by referencein their entirety.

Example 4 Specific Rotation of Corydalis Compounds

The specific rotations of several substantially pure Corydalis compoundswere determined by dissolving the compounds in ethanol and measuringtheir specific rotations using a Perkin-Elmer 241 Polarimeter. Theresults are shown in Table 2 below.

TABLE 2 Chemical Name Specific Rotation MS: m/z (M⁺ + 1)* (+)-CLMD +21370.1 14R,13S-(+)-CRDL +312 370 14R-(+)-THP +294 356 14S-(−)-THP −256356 (+)-CRPM +345 342.2

Example 5 X-Ray Diffraction of 14R,13S-Corydaline

Crystalline 14R,13S-corydaline prepared as in Example 1 was examined byX-ray diffraction as follows.

Data Collection. A Leica MZ7 polarizing microscope was used to identifya suitable specimen from a representative sampling of materials. Thechosen sample was then fixed to a nylon loop which in turn was mountedto a copper mounting pin. The mounted powder was then placed in a coldnitrogen stream (Oxford) maintained at 110K.

A BRUKER D8 GADDS general purpose three-circle X-ray diffractometer wasemployed for sample screening and data collection. The goniometer wascontrolled using the GADDS software suite (Microsoft Win 2000 operatingsystem). The sample was optically centered with the aid of a videocamera such that no translations were observed as the crystal wasrotated through all positions. The detector was set at 5.0 cm from thecrystal sample (MWPC Hi-Star Detector, 512×512 pixel). The X-rayradiation employed was generated from a Cu sealed X-ray tube(K_(α)=1.54184 Å with a potential of 40 kV and a current of 40 mA) andfiltered with a graphite monochromator in the parallel mode (175 mmcollimator with 0.5 mm pinholes).

A rotation exposure was taken to determine crystal quality and the X-raybeam intersection with the detector. The beam intersection coordinateswere compared to the configured coordinates and changes were madeaccordingly. The rotation exposure indicated acceptable crystal qualityand the unit cell determination was undertaken. Sixty data frames weretaken at widths of 0.5° with an exposure time of 10 seconds. Over 200reflections were centered and their positions were determined. Thesereflections were used in the auto-indexing procedure to determine theunit cell. A suitable cell was found and refined by nonlinear leastsquares and Bravais lattice procedures and reported here in Tables 3(14R,13S-corydaline) and 4 (14R-tetrahydropalmitine). The unit cell wasverified by examination of the hkl overlays on several frames of data,including zone photographs. No super-cell or erroneous reflections wereobserved.

After careful examination of the unit cell, a standard data collectionprocedure was initiated. This procedure consists of collection of onehemisphere of data collected using omega scans, involving the collectionover 9000 0.5° frames at fixed angles for φ, 2θ, and χ (2θ=−28°,χ=54.73°, 2θ=−90°, χ=54.73°), while varying omega. Addition data frameswere collected to complete the data set and collect Fiedel pairs. Eachframe was exposed for 10 sec. The total data collection was performedfor duration of approximately 24 hours at 110 K. No significantintensity fluctuations of equivalent reflections were observed.

After data collection, the crystal was measured carefully for size,morphology and color. Findings are reported in Table 3 and the structureis shown in FIG. 1.

TABLE 3 Crystal data and structure refinement for 14R,13S-CorydalineEmpirical formula C22H27NO4 Formula weight 369.45 Temperature 110(2) KWavelength 1.54178 Å Crystal system Monoclinic Space group P2(1) Unitcell dimensions a = 8.9648(5) Å α = 90°. b = 7.2517(4) Å β = 92.903(4)°.c = 14.9471(7) Å γ = 90°. Volume 970.46(9) Å³ Z 2 Density (calculated)1.264 Mg/m³ Absorption coefficient 0.697 mm⁻¹ F(000) 396 Crystal size0.10 × 0.05 × 0.05 mm³ Theta range for data collection 2.96 to 59.96°.Index ranges −10 <= h <= 9, −8 <= k <= 8, −16 <= 1 <= 16 Reflectionscollected 11406 Independent reflections 11406 [R(int) = 0.0000]Completeness to theta = 59.96° 99.5% Absorption correctionSemi-empirical from equivalents Max and min. transmission 0.9660 and0.9335 Refinement method Full-matrix least-squares on F²Data/restraints/parameters 11406/1/245 Goodness-of-fit on F² 1.032 FinalR indices [I > 2sigma(I)] R1 = 0.0397, wR2 = 0.0983 R indices (all data)R1 = 0.0412, wR2 = 0.1097 Absolute structure parameter 0.032(75)Extinction coefficient 0.0331(9) Largest diff. peak and hole 0.365 and−0.455 e · Å⁻³

Example 6 X-Ray Diffraction of 14R-Tetrahydropalmatine

Crystalline 14R-tetrahydropalmatine prepared as in Example 2 wasexamined by X-ray diffraction by the procedure of Example 5. Findingsare presented in Table 4 and the structure is shown in FIG. 2.

TABLE 4 Crystal data and structure refinement for14R-Tetrahydropalmitine Empirical formula C21H25NO4 Formula weight355.42 Temperature 110(2) K Wavelength 1.54178 Å Crystal systemMonoclinic Space group P2(1) Unit cell dimensions a = 15.1499(8) Å α =90°. b = 7.8440(4) Å β = 98.770(3)°. c = 15.3853(8) Å γ = 90°. Volume1806.95(16) Å³ Z 4 Density (calculated) 1.306 Mg/m³ Absorptioncoefficient 0.729 mm⁻¹ F(000) 760 Crystal size 0.10 × 0.10 × 0.01 mm³Theta range for data collection 2.91 to 59.97°. Index ranges −16 <= h <=17, −8 <= k <= 8, −17 <= 1 <= 17 Reflections collected 13614 Independentreflections 13614 [R(int) = 0.0000] Completeness to theta = 59.97° 99.8%Absorption correction Semi-empirical from equivalents Max and min.transmission 0.9927 and 0.9307 Refinement method Full-matrixleast-squares on F² Data/restraints/parameters 13614/1/477Goodness-of-fit on F² 1.099 Final R indices [I > 2sigma(I)] R1 = 0.0430,wR2 = 0.0984 R indices (all data) R1 = 0.0627, wR2 = 0.1282 Absolutestructure parameter 0.00(13) Largest diff. peak and hole 0.201 and−0.183 e · Å⁻³

Example 7 Synthesis, Extraction and/or Plant Sources

Exemplary syntheses and identification related to isolation of acompound of the invention are also generally described by Boudou et al.,J. Org. Chem., 70, 9486-94 (2005), and Shaath et al., J. Org. Chem., 40,1987-88 (1975), each of which are incorporated by reference herein.

Example 8 Design and Synthesis of Compounds

1. Preparation of Berberrubine from Berberine.

Berberine (1.0 g, 2.68 mmol) was heated at 190° in a dry oven undervacuum for 30 minutes. The crude product was recrystallized from EtOH togive berberrubine (0.6 g, yield 60%, confirmed by ¹HNMR).

2. Preparation of Compound I1 from Berberrubine.

A suspension of berberrubine chloride (0.2 g, 0.5 mmol) and1,3-dibromopropane (0.58 g, 2.8 mmol) in dry DMF was heat at 60°. Thesuspension was cooled to room temperature and ethyl ether was added. Theprecipitate was collected by filtration, rinsed with ethyl ether anddried under vacuum to give compound I1 as yellow solid (confirmed by ¹HNMR). The reaction was rerun on 0.5 g scale (berberrubine), giving 0.55g of crude compound I1 (yield 82%).

3. Preparation of Compound 67 from Corypalmine

D-Biotin (105 mg, 0.43 mmol), EDC HCl (125 mg, 0.65 mmol) and DMAP (19mg, 0.16 mmol) were dissolved together in a flask with a minimum volumeof DMF (4.5 mL). Then corypalmine (25 mg, 0.073 mmol) was dissolved inthis solution. After stirring for 3 hours, 0.5 mL sample of the reactionsolution was taken out for testing. The sample was added to 10 mL H₂Oand extracted with 10 mL EtOAc. The organic layer was dried with MgSO₄,filtered, and concentrated under vacuum. The product was subjected toLC-MS. The LC-MS information suggested formation of compound 67. Theremaining reaction mixture was stirred overnight. The reaction mixturewas subsequently extracted with EtOAc. The organic layer was dried withMgSO₄ and evaporated in vacuo. The yellow residue was isolated bypreparative TLC to afford compound 67. Preparative HPLC was used topurify compound, 67. The purified product was confirmed by MS and HPLCto confirm the structure and the purity (92.1% and 93.1%).

4. Preparation of Compound I2 from Corypalmine.

Corypalmine (0.068 g, 0.2 mmol) was added to 20 mL acetone and 5 mLethanol. The suspension was refluxed for 1 hour to dissolve the startingmaterial. Then 0.068 mg K₂CO₃ was added and the suspension was refluxedfor 1 hour. Ethyl 2-bromoacetate (0.0244 mL, 0.22 mmol) was dissolved in1 mL acetone and added into the reaction suspension in portions over 30minutes. The resulting suspension was refluxed for 2 hours. The reactionwas monitored by LC-MS. Part of the product was purified by preparativeTLC.

5. Preparation of Compound 68 from Compound I2.

Compound I2, prepared according to procedure 4 was used withoutpurification, and saponified with NaOH to prepare compound 68. Thisreaction was monitored by LC-MS. After standard workup.

6. Preparation for Compound I3 from Berberrubine.

Berberrubine (60 mg, 0.1 mmol) was added to 7 mL hot MeOH and stirredfor 15 minutes at 60° C. Then NaBH₄ (8 mg, 0.21 mmol) was added to themixture and the mixture was stirred at 60° for 15 minutes. Five mL H₂Owas added to the solution to quench the reaction. The product 13 wasextracted from the solution with CHCl₃ (10 mL×3). 10 mg grey solid wasobtained, yield 20%. The isolated material gave the expected peak in MSand confirmed the structure. HPLC analysis suggested the purity wassatisfactory for use without further purification.

7. Preparation for Compound 74 from Compound I1

Compound I1 (80 mg, 0.168 mmol) was dissolved in CH₃OH (5 mL) in a 25 mLflask at rt. The color of the solution was dark red. Sodium borohydride(8 mg, 0.210 mmol) was added to this flask. The reaction solution becamelight yellow soon thereafter. The reaction was maintained at rf for 2hours. Then H₂O (20 mL) was added to the solution to quench thereaction.

The solvent was evaporated under reduced pressure to remove all of theCH₃OH. Then 30 mL water were added to the residue and the solution wasextracted with chloroform 3 times. The combined organic extracts weredried over anhydrous MgSO₄, filtered, and concentrated under vacuum. 40mg product was obtained as yellow oil. (yield: 54.4%). The MS analysisconfirmed the structure of compound 74. The HPLC analysis suggested thepurity was 86%.

8. Preparation for Compound 77 from 14R-(+)-THP.

14R-(+)-THP (250 mg, 0.70 mmol) was added to 15 mL 47% HBr and stirredovernight at 100° C. Then the solution was cooled to room temperature,and the product was filtered off to give compound 77 as the hydrobromidesalt.

9. Preparation for Compound 78 from 14R,13S-(+)-CDRL.

14R,13S-CDRL (200 mg, 0.54 mmol) was added to 10 mL 47% HBr and stirredovernight at 100° C. Then the solution was cooled to rt, and the productwas filtered off to give compound 78 as the hydrobromide salt.

10. Preparation for Compound 79 from Berberrubine.

Berberrubine (248 mg, 0.693 mmol) was added to 5 mL CHCl₃ in a flask.The mixture was stirred and refluxed. Then benzenesulfonyl chloride (760mg, 4.30 mmol) and pyridine (0.1 mL) were slowly added in. The reactionmixture was stirred at the same temperature for 2 hours, andsubsequently cooled down to room temperature. The mixture was filteredto afford a yellow solid. The solid was washed with CHCl₃ 3 times anddried under vacuum to provide the final product, compound 79, (yellowsolid, 204 mg). Compound 79 was used directly for the next step withoutpurification.

11. Preparation for Compound 69 from Compound 79.

Compound 79 (132 mg, 0.265 mmol) was added in a flask with methanol (5mL). Then the reaction mixture was heated to reflux to dissolve thestarting material. Then NaBH₄ (42 mg, 1.11 mmol) was added slowly to theflask. The reaction mixture was stirred at the same temperature for 1hour. Then it was cooled down to room temperature and then cooled inrefrigerator for 4 hours. The mixture was filtered to afford lightyellow crystals. The crystals were washed with H₂O 3 times and thendried under vacuum to provide the final product compound 69 (lightyellow solid, 28.7 mg). NMR & MS analyses were consistent with thestructure of compound 69.

Analogs of compound 69 may be readily made using commercially availablesubstituted phenyl sulfonyl chlorides.

12. Preparation for Compound 80 from Berberrubine.

Berberrubine (0.5 g, 1.4 mmol) was dissolved in 40 mL CHCl₃ byrefluxing. After stirring for about 30 minutes, the methanesulfonylchloride (0.33 mL, 4.2 mmol) was added to the solution dropwise over 30seconds. 10 minutes later, yellow solid appeared. The suspension wasrefluxed for 3 hours. After cooling, the suspension was filtered toprovide a yellow solid. This intermediate (compound 80) was useddirectly for the next step without purification.

13. Preparation for Compound 71 from Compound 80.

Compound 80 was dissolved in 30 mL MeOH at reflux. NaBH₄ (0.052 g, 1.3mmol) was added to the reaction. The reaction occurred immediately. Thesolution was refluxed for 2 hours and cooled down. Analysis by TLCsuggested the transformation was complete. The reaction was leftovernight, and a gray solid appeared the next morning. The suspensionwas filtered to afford the gray solid. NMR & MS were consistent with thestructure of the compound 71.

14. Preparation for Compound 81 from Berberrubine.

Berberrubine (300 mg, 0.84 mmol) was added to 15 mL CHCl₃ and stirredfor 20 minutes at reflux. Benzoyl chloride (1180 mg, 8.4 mmol) and 0.1mL pyridine were added to the solution. The mixture was stirred for 2hours at reflux. Then product was filtered, washing with CHCl₃. Thestructure of compound 81w as confirmed by ¹H NMR and MS.

15. Preparation for Compound 70 from Compound 81.

The reaction mixture of the previous reaction was continued by additionof NaBH₄ (8 mg, 0.21 mmol). Then the mixture stirred at 60° C. for 15minutes. 5 mL H₂O was added to the solution to quench the reaction. Theproduct was filtered from the solution. MS information suggested it wasthe desired structure.

16. Preparation for Compound I4 from Berberrubine.

Berberrubine (100 mg, 0.279 mmol) was dissolved in 5 mL acetone. Then2-bromoethanol (182 mg, 1.40 mmol) was added to the solution and stirredovernight at 60°. The anticipated yellow solid was appeared. Then theproduct was filtered from the reaction and used without furtherpurification.

17. Preparation for Compound 82 from Compound I4.

Compound I4 (60 mg, 0.186 mmol) was added to 5 mL hot MeOH and stirredfor 15 minutes at 60°. Then NaBH₄ (9 mg, 0.24 mmol) was added to thesolution. The color of the solution changed immediately. Then themixture stirred at 60° for 15 minutes, then worked up and isolated asbefore.

18. Preparation for Compound I5 from Berberrubine.

Berberrubine (400 mg, 1.12 mmol) was dissolved in CHCl₃ (14 mL) atreflux in a 50 mL flask. Then ethyl bromoacetate (1.5 g, 8.96 mmol) wasadded dropwise to the reaction. After the ethyl bromoacetate had beenadded entirely, the red solution changed into yellow. The reaction wasmaintained at reflux.

The solution was filtered and the solid was refluxed in CHCl₃ (15 mL)for 2 hours and filtered. ¹H NMR information suggested the product wasthe desired structure 15.

19. Preparation for Compound 83 from Compound I5.

Compound I5 (250 mg, 0.563 mmol) was placed in 50 mL flask and CH₃OH (12mL) was added in. The solution became clear after refluxing for a while.Then sodium boro-hydride (27.7 mg, 0.732 mmol) was added carefully intothe same flask. TLC showed that the material has been disappeared. Thereaction continued and was held at the same temperature for 40 minutes.

Water (30 mL) was added to the solution, stirred for half an hour. Thenthe solution was evaporated under vacuum until the CH₃OH was removed.Water (30 mL) was added into the solution, and the solution wasextracted by chloroform for 3 times. The organic phase was washed bywater and brine, the dried by anhydrous sodium sulfate, evaporated undervacuum to give compound 83.

20. Preparation for Compound I6 from Berberrubine.

Berberrubine (648 mg, 1.81 mmol) was added in a flask with 15 mLN,N-dimethylformamide in it. The mixture was stirred and heated todissolve under 60° C. Then 2-bromoacetic acid (724 mg, 5.21 mmol) wasslowly added in. The reaction mixture was stirred under the sametemperature for 2 hours. Then it was cooled down to room temperature.The mixture was filtered to show the yellow solid. Then the solid waswash with CHCl₃ for 3 times and then dried in vacuum to show theproduct.

21. Preparation for Compound III from Compound I6.

The compound I6 (109 mg, 0.262 mmol) was added in a flask with methanol(5 mL). Then the reaction mixture was heated to dissolve under refluxconditions. NaBH₄ (42 mg, 1.11 mmol) was added to the reaction slowly.The reaction mixture was stirred at the same temperature for 1 hour.Then it was cooled first to room temperature and then cooled in arefrigerator for 4 hours. The mixture was filtered to give clearcrystals. The crystals were washed with Et₂O for 3 times and dried invacuum to give the final product, compound 111.

22. Preparation for Compound I7 from Berberrubine.

Berberrubine (1.0 g, 2.79 mmol) was dissolved in 5 mL DMF. Then1,2-dibromoethane (5.3 g, 27.9 mmol) was added to the solution andstirred overnight at 60° C. Then 5 mL Et₂O was added to the solution.The product was filtered from the reaction and used directly for thenext reaction.

23. Preparation for Compound I8 from Compound I7.

A solution of compound I7 (460 mg, 1 mmol), morpholine (870 mg, 10mmol), K₂CO₃ (1.38 g, 10 mmol), DMF (30 mL) was heated at 60° C.

24. Preparation for Compound 88 from Compound I8.

Compound I8 was dissolved in 50 mL methanol and heated to reflux. NaBH₄was added to the refluxing solution. The reaction was monitored by TLCand was refluxed for 4 hours. The methanol was removed by vacuumdistillation. The resulting residue was mixed with water and extractedwith chloroform (3×). The organic layer was dried over NaSO₄. Then thechloroform was removed by vacuum distillation. The product was purifiedby silica gel column chromatography (acetone: CH₂Cl₂, from 4:1 to 1:1).The MS analysis was consistent with the desired product and the HPLCassay showed the purity was 96%. ¹H NMR confirmed the structure was thedesired one, compound 88.

25. Preparation for Compound 72 from Compound 78.

A solution of compound 78 (100 mg, 0.32 mmol) in 5 mL dry DMF under Arwas heated to 60° C. and a solution of CH₂BrCl (84 mg, 0.64 mmol) wasadded. The temperature of the reaction was raised to 95° C. 4 hourslater, the reaction was analyzed by LC-MS and confirmed that the productwas compound 72.

26. Preparation of Compound I9 from Berberrubine.

Berberrubine (300 mg, 0.84 mmol) was dissolved in 30 mL CHCl₃ at reflux.Then 4-(trifluoromethyl)benzene-1-sulfonyl chloride (300 mg, 1.23 mmol)was added to the solution and stirred for 7 hours. Then product wasfiltered, washed with CHCl₃ and dried. 400 mg product was obtained asyellow solid (yield: 84.2%). The intermediate was used directly withoutfurther purification.

27. Preparation of Compound 85 from Compound I9.

Compound I9 (140 mg, 0.247 mmol) was dissolved in 80 mL MeOH at reflux.NaBH₄ (50 mg, 1.322 mmol) was added to the solution and stirred for 1hour. Then product was filtered, washed with MeOH, H₂O and hexane. 45 mgproduct was obtained as gray solid (yield: 34%). The MS & ¹H NMRanalysis of the product was consistent with the structure of compound85.

28. Preparation of Compound I10 from Berberrubine.

Berberrubine (300 mg, 0.84 mmol) was dissolved in 30 mL CHCl₃ at 70° C.Then 3,4-dimethoxybenzene-1-sulfonyl chloride (300 mg, 1.27 mmol) wasadded to the solution and stirred for 7 hours. Then product wasfiltered, washed by CHCl₃ and dried. 40 mg product was obtained asyellow solid. (yield: 17%). The intermediate was used directly withoutfurther purification.

29. Preparation of Compound 86 from Compound I10.

Compound I10 (80 mg, 0.14 mmol) was dissolved in 80 mL MeOH at 70° C.NaBH₄ (50 mg, 1.32 mmol) was added to the solution and stirred for 1hours. Then product was filtered, washed with MeOH, H₂O and hexane. 25mg product was obtained as gray solid. (MC0236-38-1; yield: 34%). The MS& ¹H NMR information suggested the product was the desired structure,compound 86.

30. Preparation of Compound I11 from Berberrubine.

Berberrubine (300 mg, 0.84 mmol) was dissolved in 30 mL CHCl₃ at 70° C.Then 4-nitrobenzene-1-sulfonyl chloride (300 mg, 1.35 mmol) was added tothe solution and stirred for 7 hours. Then product was filtered, washedby CHCl₃ and dried. 400 mg product was obtained as yellow solid. (yield:87.7%) The intermediate was used directly without further purification.

31. Preparation of Compound 106 from Compound I11.

Compound I11 (110 mg, 0.202 mmol) was dissolved in CH₃OH (150 mL) atreflux in a 250 mL three-neck flask. Then sodium borohydride (100 mg,2.64 mmol) was added carefully to the reaction. The mixture was refluxedfor 3 hours. The solution was concentrated to 10 mL in vacuum, cooled inrefrigerator, filtered, washed by water and n-hexane, dried in vacuum.35 mg product was obtained as cyan solid. (yield: 33.9%)

32. Preparation of Compound 107 from Compound I12.

Compound I12 (130 mg, 0.325 mmol) was dissolved in CH₃OH (150 mL) atreflux in a 250 mL three-neck flask. Then sodium borohydride (120 mg,3.17 mmol) was added carefully. The reaction continued for 3 hours. Thesolution was concentrated to 10 mL in vacuum, cooled in refrigerator,filtered, washed by water and n-hexane, dried in vacuum.

33. Preparation of Compound I13 from Berberrubine.

Berberrubine (200 mg, 0.56 mmol) was dissolved in 30 mL CHCl₃ at 70° C.Then 4-(methylsulfonyl)benzene-1-sulfonyl chloride (350 mg, 1.37 mmol)was added to the solution and stirred for 7 hours at 70° C. Then productwas filtered, washed by CHCl₃ and dried. 200 mg product was obtained asyellow solid. (yield: 62%) The intermediate compound I13 was useddirectly without further purification.

34. Preparation of Compound 87 from Compound I13.

Compound I13 (110 mg, 0.19 mmol) was dissolved in 80 mL MeOH at reflux.Then NaBH₄ (70 mg, 1.85 mmol) was added to the solution and stirred for1 hour at 70° C. Then most of solvent was removed, and product wasfiltered, washed by MeOH, H₂O and hexane. 40 mg product was obtained asgray solid (yield: 38.8%).

35. Preparation of Compound I14 from Berberrubine.

Berberrubine (220 mg, 0.61 mmol) was dissolved in 30 mL CHCl₃ at 70° C.Then 4-cyanobenzene-1-sulfonyl chloride (340 mg, 1.68 mmol) was added tothe solution and stirred for 7 hours at 70° C. Then product wasfiltered, washed by CHCl₃ and dried. 110 mg product was obtained asyellow solid. (yield: 35%) The intermediate compound I14 was useddirectly without further purification.

36. Preparation of Compound 108 from Compound I14.

37. Preparation of Compound 109 from Compound I15.

The Compound I15 (150 mg, 0.449 mmol) was added in a flask with methanol(15 mL). Then the reaction mixture was heated to dissolve under refluxcondition. Then NaBH₄ (276 mg, 7.30 mmol) was added in slowly. Thereaction mixture was stirred under the same temperature for 2 hours.Then it was evaporated in vacuum and the solid was washed with water for24 hours. The mixture was filtered and then dried to obtain 112 mgcompound 109 as red solid. (Yield: 81.2%) The ¹H NMR & MS informationsuggested the product was the desired structure.

38. Preparation of Compound 110 from Compound 109.

Compound 109 (17 mg, 0.0461 mmol) was added to a flask with 13 mL CHCl₃in it. The mixture was stirred and heated to reflux to dissolve it. Thenbenzenesulfonyl chloride (55 mg, 0.311 mmol) and pyridine (0.3 mL) wereslowly added in. The reaction mixture was stirred at the sametemperature for 2 hours. Then the reaction was cooled down to roomtemperature. The MS assay suggested the compound 110 was formed.

39. Preparation of Compound I17 from Compound I16.

A solution of thiophenol (229 mg, 1.8 mmol) in 5 mL toluene was treatedwith DBU (317 mg, 1.8 mmol). 5 minutes later, 15 mL of toluene solution,which contained compound I16 (300 mg, 0.6 mmol), was added to thereaction solution. The reaction was stirred overnight. The LC-MSinformation suggested the desired product compound I17 was formed. Thenproduct was purified by silica gel column chromatogram. (300 mL CH₂Cl₂,then 300 mL CH₂Cl₂: acetone=3:1). 200 mg product was obtained. (Yield:62.3%) The ¹H NMR & MS information suggested the product was the desiredstructure.

40. Preparation of Compound 105 from Compound I17.

The compound I17 (80 mg, 0.17 mmol) was dissolved in 5 mL AcOH andheated at 60° C. 10 minutes later, the 2 mL 30% H₂O₂ was added in. 1.5hours later, the reaction was sent to LC-MS assay.

41. Preparation of Compound 1.

Berberine Chloride→B

To a stirred solution of NaOH (4 g, 100 mmol) in water (20 mL) was addedberberine chloride (2.0 g, 5.4 mmol) at room temperature. Then acetone(1.6 mL) was added slowly at same temperature and stirred for 1 hour.TLC analysis indicated the completion of the reaction. The reactionsolution was filtered, sufficiently washed with 80% methanol and driedto give 1.7 g of B.

B→C

To a stirred solution of B (1.7 g, 4.3 mmol) in MeCN (20 mL) was addedKI (450 mg, 2.7 mmol) at room temperature. The reaction mixture washeated to reflux and BnBr (1.5 mL, 12.7 mmol) was added. Then thereaction was refluxed for 4 hours with stirring. TLC analysis indicatedthe completion of the reaction. The solvent was removed and residue waspurified by column chromatography to give 1.6 g of C.

C→Compound 1

To a stirred solution of C (1.6 g, 3.7 mmol) in MeOH was added Pt/C (200mg) and stirred overnight under H₂ at ambient temperature. Afterfiltering, the filtrate was concentrated to get crude product. The solidwas washed with MeOH to get 500 mg of pure Compound 1. ¹H NMR (CDCl₃): δ7.20-7.14 (m, 3H); 6.88-6.85 (m, 2H); 6.76 (s, 1H); 6.61 (s, 1H); 6.52(d, 1H, J=8.4 Hz); 6.00 (d, 1H, J=8.4 Hz); 5.94 (s, 2H); 4.28 (d, 1H,J=16.2 Hz); 3.86 (s, 3H); 3.80 (s, 3H); 3.77 (br, 1H); 3.56 (d, 1H,J=16.2 Hz); 3.25-3.07 (m, 3H); 2.73-2.57 (m, 4H). MS: m/z (APC)-ESI)430.2 (M⁺+1).

42. Preparation of Compounds, Salt Formation.

General Procedure: 14R,13S-(+)-CRDL or 14R-(+)-THP was dissolved insolvent and mixed with a suitable amount of acid to form thecorresponding acid addition salt.

Hydrochloride Salts (Compounds 2 and 6)

To a stirred solution of MeOH (2 mL) and DCM (2 mL) contained HCl (2mmol) was added 14R-(+)-THP (100 mg, 0.28 mmol) or 14R,13S-(+)-CDRL (100mg, 0.27 mmol) at room temperature and stirred for 2 hours. The solventwas removed to get 110 mg of the salt.

Sulfate Salts (Compounds 3 and 7)

To a stirred solution of MeOH (2 mL) and DCM (2 mL) contained H₂SO₄ (15μL, 0.28 mmol) was added 14R-(+)-THP (100 mg, 0.28 mmol) or14R,13S-(+)-CDRL (100 mg, 0.27 mmol) at room temperature and stirred for2 hours. The solvent was removed to get 110 mg of the salt.

Citrate Salts (Compounds 4 and 8)

To a stirred solution of MeOH (2 mL) and DCM (2 mL) contained citricacid (19.6 mg, 0.093 mmol) was added 14R-(+)-THP (100 mg, 0.28 mmol) or14R,13S-(+)-CDRL (100 mg, 0.27 mmol) at room temperature and stirred for2 hours. The solvent was removed to get 110 mg of the salt.

Maleate Salts (Compounds 5 and 9)

To a stirred solution of MeOH (2 mL) and DCM (2 mL) contained maleicacid (16.2 mg, 0.14 mmol) was added 14R-(+)-THP (100 mg, 0.28 mmol) or14R,13S-(+)-CDRL (100 mg, 0.27 mmol) at room temperature and stirred for2 hours. The solvent was removed to get 110 mg of the salt.

43. Preparation of Compound 10.

A+B→C

To the solution of A (650 mg, 3.31 mmol), B (630 mg, 3.48 mmol) in DCM(30 mL) was added EDCI (1.27 g, 6.62 mmol) and Et₃N (1.0 g, 9.93 mmol)at 20° C. and the solution was stirred overnight. TLC analysis indicatedthe completion of the reaction. Water was added, and the organic layerwas collected, dried and concentrated to give 1.0 g of C.

C→D

To a stirred solution of C (1.0 g, 2.78 mmol) in toluene (10 mL) wasadded POCl₃ (3 mL) at ambient temperature. The reaction mixture wasrefluxed for 4 hours with stirring. When the reaction was completed, thesolvent and excess POCl₃ were evaporated off. The residue was pouredinto ice water and adjusted pH>7 with Na₂CO₃. The solution was extractedwith EtOAc, and the organic layer was dried and concentrated to get thecrude product, which was directly used in the next step without furtherpurification.

D→E

To a stirred solution of D in MeOH (15 mL) was added NaBH₄ (130 mg, 3.42mmol) at 10° C. The reaction mixture was stirred for 6 hours at roomtemperature. TLC analysis indicated the completion of the reaction. Thesolvent was removed and the residue was extracted with EtOAc and water.The organic layer was dried and concentrated to give crude E.

E→Compound 10

Crude E was added to 37% HCHO (5 mL) and AcOH (10 mL). The reactionmixture was heated to 100° C. and stirred for 8 hours. TLC analysisindicated the completion of the reaction. The water and AcOH wasremoved. The residue was extracted with Na₂CO₃ aqueous and EtOAc. Theorganic phase was dried and concentrated to crude product. The crudematerial was further purified by column chromatography to get 200 mg ofcompound 10. ¹H NMR (CDCl₃): δ 6.74 (s, 1H); 6.67 (s, 1H); 6.62 (s, 1H);6.58 (s, 1H); 3.97-3.92 (m, 1H); 3.89-3.85 (m, 12H); 3.71 (s, 1H);3.65-3.56 (m, 1H); 3.27-3.22 (m, 1H); 3.18-3.14 (m, 2H); 2.87-2.78 (m,1H); 2.68-2.63 (m, 2H). MS: m/z (APC)-ESI) 356.1 (M⁺+1).

44. Preparation of Compound 11.

A+B→C

To the solution of A (1.8 g, 10 mmol), B (1.81 g, 10 mmol) in DCM (20mL) was added EDCI (2.83 g, 12 mmol) and Et₃N (1.0 g, 9.93 mmol) at 20°C. and the solution was stirred overnight. TLC analysis indicated thecompletion of the reaction. Water was added, and the organic layer wascollected, dried and concentrated to give 1.9 g of C.

C→D

To a stirred solution of C (0.3 g, 0.88 mmol) in toluene (10 mL) wasadded POCl₃ (1.5 mL) at ambient temperature. The reaction mixture wasrefluxed for 4 hours with stirring. When the reaction was completed, thesolvent and excess POCl₃ were evaporated off. The residue was pouredinto ice water and adjusted pH>7 with Na₂CO₃. The solution was extractedwith EtOAc, and the organic layer was dried and concentrated to get thecrude product, which was directly used in the next step without furtherpurification.

D→E

To a stirred solution of D in MeOH (8 mL) was added NaBH₄ (130 mg, 3.42mmol) at 10° C. The reaction mixture was stirred for 6 hours at roomtemperature. TLC analysis indicated the completion of the reaction. Thesolvent was removed and the residue was extracted with EtOAc and water.The organic layer was dried and concentrated to give crude E (123 mg).

E→Compound 11

Crude E was added to 37% HCHO (10 mL) and AcOH (10 mL). The reactionmixture was heated to 100° C. and stirred for 8 hours. TLC analysisindicated the completion of the reaction. The water and AcOH wasremoved. The residue was extracted with Na₂CO₃ aqueous and EtOAc. Theorganic phase was dried and concentrated to crude product. The crudematerial was further purified by column chromatography to get 78 mg ofcompound 11. MS (M+1): 340.1. ¹H NMR (CDCl₃) δ=6.73 (s, 1H), 6.61-6.63(s, 2H), 6.55 (s, 1H), 6.70-5.91 (s, 2H), 3.89-3.94 (m, 1H), 3.87-3.90(s, 6H), 3.54-3.670 (m, 2H), 3.10-3.25 (m, 3H), 2.80-2.86 (t, 1H),2.60-2.70 (m, 2H).

45. Preparation of Compounds 12 and 13.

A→B

To the solution of A (1.0 g, 2.95 mmol) in dry THF was added n-BuLi (2.5M, 3.25 mmol) dropwise at the protection of N₂ under the temperature of−30° C., stirring for 1 hour, then CH₃CH₂Br (386 mg, 3.54 mmol) wasadded dropwise. After the addition was finished, the solution wasstirring at room temperature for 1.5 hours. NH₄Cl was added and theresolution was removed in vacuo. Water was added and the product wasextracted with CH₂Cl₂, purified by flash chromatography to afford 968mg, yield: 89.5%.

B→C

To the solution of B (500 mg) in AcOH was added PtO₂ (45 mg), stirringovernight under an atmosphere of H₂ (8 atm) at 25° C. When the reactionwas finished, the solution was concentrated in vacuo. Then water wasadded, and the solution was adjusted to PH=7 using Na₂CO₃. The solutionwas extracted with CH₂Cl₂ and the crude product was purified by flashchromatography to afford 300 mg of product, yield: 59.3%.

C→Compounds 12 and 13

C (300 mg) was dissolved in the solution of HCHO (37%, 20 mL) and AcOH(20 mL). The reaction mixture was heated to reflux for 6 hours and thenconcentrated in vacuo and extracted by CH₂Cl₂. The residue was purifiedby flash chromatography and TLC to afford two products, one is compound12 (15.6 mg), and another is compound 13 (19.6 mg).

Compound 12. ¹H NMR (CDCl₃) δ=6.71 (s, 1H), 6.66 (s, 1H), 6.60 (s, 1H),6.51 (s, 1H), 4.41-4.05 (m, 1H), 4.01-3.95 (m, 1H), 3.85 (s, 6H), 3.83(s, 6H), 3.76-3.71 (m, 1H), 3.10-3.05 (m, 1H), 2.95-2.94 (m, 2H),2.10-1.90 (m, 2H), 1.24 (m, 1H), 0.95-0.90 (t, 3H). MS: m/z (APCI-ESI):384.2 (M⁺+1).

Compound 13. ¹H NMR (CDCl₃) δ=6.70 (s, 1H), 6.68 (s, 1H), 6.60 (s, 1H),6.58 (s, 1H), 4.40-3.98 (m, 1H), 3.88 (s, 6H), 3.85 (s, 6H), 3.74 (s,1H), 3.10-3.08 (m, 1H), 3.10-3.08 (m, 1H), 2.98-2.92 (m, 1H), 2.60-2.54(m, 2H), 1.42-1.37 (m, 2H), 0.83-0.78 (t, 3H). MS: m/z (APCI-ESI): 384.2(M⁺+1).

46. Preparation of Compounds 14 and 15.

To a solution of 210 mg (1.10 mmol) of A in 10 mL of toluene, 180 mg(1.11 mmol) of B was added and the mixture was heated at reflux for 2hours. The mixture was allowed to cool to room temperature and was leftto stand overnight. The crystals formed was collected, washed withdiethyl ether and dried under vacuum. 350 mg of crude product 14 wasobtained. ¹H NMR (300 MHz, DMSO-d₆) δ=7.88 (d, J=8.7 Hz, 1H), 7.56-7.51(m, 1H), 7.41-7.39 (m, 2H), 6.94 (s, 1H), 6.75 (s, 1H), 5.27 (d, J=6.0Hz, 1H), 4.6-4.58 (m, 2H), 3.69 (s, 3H), 3.68 (s, 3H), 3.18-3.09 (m,1H), 3.01-2.90 (m, 1H), 2.72-2.67 (br, 1H). MS: m/z (APCI-ESI): 354.1(M⁺+1).

A solution of 200 mg (0.57 mmol) of compound 14 in AcOH was heated atreflux for 24 hours. The solvent was evaporated, and the residue waspurified by washed with PE and EtOAc to yield 90 mg of compound 15. ¹HNMR (300 MHz, DMSO-d₆) δ=7.97 (d, J=7.6 Hz, 1H), 7.59-7.43 (m, 3H), 7.12(s, 1H), 6.78 (s, 1H), 5.19 (d, J=4.4 Hz), 4.80 (d, J=5.7 Hz), 4.45 (d,J=4.5 Hz), 3.77 (s, 3H), 3.75 (s, 3H), 2.90-2.66 (m, 3H). MS: m/z(APCI-ESI): 354.1 (M⁺+1).

47. Preparation of Compound 16.

A+B→C

To the solution of A (1.82 g, 10 mmol), B (1.81 g, 10 mmol) in DCM (30mL) was added EDCI (2.83 g, 12 mmol) and Et₃N (1.0 g, 9.93 mmol) at 20°C. and the solution was stirred overnight. TLC analysis indicated thecompletion of the reaction. Water was added, and the organic layer wascollected, dried and concentrated to get 2.85 g of C.

C→D

To a stirred solution of C (2.55 g, 7.2 mmol) in toluene (20 mL) wasadded POCl₃ (8 mL) at ambient temperature. The reaction mixture wasrefluxed for 4 hours with stirring. When the reaction was completed, thesolvent and excess POCl₃ were evaporated off. The residue was pouredinto ice water and adjusted pH>7 with Na₂CO₃. The solution was extractedwith EtOAc, and the organic layer was dried and concentrated to get thecrude product, which was directly used in the next step without furtherpurification.

D→E

To a stirred solution of D in MeOH (30 mL) was added NaBH₄ (1.0 g) at10° C. The reaction mixture was stirred for 6 hours at room temperature.TLC analysis indicated the completion of the reaction. The solvent wasremoved and the residue was extracted with EtOAc and water. The organiclayer was dried and concentrated to get crude E (1.3 g).

E→Compound 16

Crude E (0.6 g) was added to 37% HCHO (15 mL) and AcOH (15 mL). Thereaction mixture was heated to 100° C. and stirred for 8 hours. TLCanalysis indicated the completion of the reaction. The water and AcOHwas removed. The residue was extracted with Na₂CO₃ aqueous and EtOAc.The organic phase was dried and concentrated to crude product. The crudematerial was further purified by column chromatography to get 160 mg ofcompound 16. MS (M+1): 342.1. ¹HNMR (CDCl₃) δ (ppm) 6.64 (s, 1H),6.62-6.63 (s, 2H), 6.73 (s, 1H), 3.89-3.94 (m, 1H), 3.86-3.94 (s, 3×3H),3.55-3.66 (m, 2H), 3.11-3.27 (m, 3H), 2.84-2.88 (t, 1H), 2.60-2.69 (m,2H).

48. Preparation of Compound 17.

A+B→C

To the solution of A (1.82 g, 10 mmol), B (1.81 g, 10 mmol) in DCM (30mL) was added EDCI (2.83 g, 12 mmol) and Et₃N (1.0 g, 9.93 mmol) at 20°C. and the solution was stirred overnight. TLC analysis indicated thecompletion of the reaction. Water was added, and the organic layer wascollected, dried and concentrated to get 2.3 g of C.

C→D

To a stirred solution of C (0.25 g, 0.72 mmol) in toluene (10 mL) wasadded POCl₃ (1.5 mL) at ambient temperature. The reaction mixture wasrefluxed for 4 hours with stirring. When the reaction was completed, thesolvent and excess POCl₃ were evaporated off. The residue was pouredinto ice water and adjusted pH>7 with Na₂CO₃. The solution was extractedwith EtOAc, and the organic layer was dried and concentrated to get thecrude product, which was directly used in the next step without furtherpurification.

D→E

To a stirred solution of D in MeOH (8 mL) was added NaBH₄ (130 mg, 3.42mmol) at 10° C. The reaction mixture was stirred for 6 hours at roomtemperature. TLC analysis indicated the completion of the reaction. Thesolvent was removed and the residue was extracted with EtOAc and water.The organic layer was dried and concentrated to get crude E (140 mg).

E→Compound 17

Crude E (120 mg) was added to 37% HCHO (5 mL) and AcOH (5 mL). Thereaction mixture was heated to 100° C. and stirred for 8 hours. TLCanalysis indicated the completion of the reaction. The water and AcOHwas removed. The residue was extracted with Na₂CO₃ aqueous and EtOAc.The organic phase was dried and concentrated to crude product. The crudematerial was further purified by column chromatography to get 200 mg ofcompound 17. MS (M+1): 342.1. ¹H NMR (CDCl₃) δ (ppm) 6.72-6.73 (s, 2H),6.611 (s, 1H), 6.554 (s, 1H), 3.89-3.94 (m, 1H), 3.87-3.90 (s, 3×3H),3.55-3.69 (m, 2H), 3.11-3.24 (m, 3H), 2.79-2.84 (t, 1H), 2.60-2.68 (m,2H).

49. Preparation of Compounds 19 and 20.

A+B→C

A mixture of 200 mg (1.10 mmol) of A and 165 mg (1.10 mmol) of B washeated to 160° C. for 1 hour, and then cooled; the oil was dissolved in2 mL of MeOH. 50 mg (1.32 mmol) of NaBH₄ was added in portions. After 1hour at room temperature, the reaction was completed, and the solventwas removed under vacuum. The residue was dissolved with ethyl acetate,washed with water, brine, and dried over anhydrous sodium sulfate. 320mg of pure product was obtained. Yield: 92.4%.

C→Compound 19

To a solution of 150 mg (0.48 mmol) of C in 2 mL of AcOH was added 1.0 gof CuSO₄, followed 2 mL of aqueous (30%) glyoxal. The reaction mixturewas heated to reflux for 3 hours. The most solvent was removed undervacuum, and then extracted with EtOAc. The organic phase was washed withwater, brine and dried over anhydrous NaSO₄. The solvent was removedunder vacuum, the crude product was purified with column chromatographeluting with EA:MeOH=20:1.35 mg of the title compound was obtained.Yield: 21.0%. ¹H NMR (CDCl₃, 300 MHz) δ=8.81 (s, 1H), 8.41 (br, 1H),7.99 (d, J=8.7 Hz), 7.51 (d, J=8.7 Hz), 6.94 (s, 1H), 6.35 (s, 2H), 4.66(t, J=5.7 Hz, 2H), 3.81 (s, 3H), 3.78 (s, 3H), 3.06 (t, J=6.2 Hz, 2H).MS: m/z (APCI-ESI): 352.1 (M⁺+1).

Compound 19→Compound 20

To a suspension of 14 mg (0.04 mmol) of compound 19 in 2 mL of1,2-dichloroethane was added 1 mL of POCl₃. The mixture was stirred for30 minutes at 80° C., and then the solvent was removed in vacuo. Theresidue was dissolved in MeOH, then NaBH₄ was added in portions at 0° C.After 30 minutes, the reaction was completed. The solvent was removedunder vacuum, the residue was extracted with CH₂Cl₂, washed with water,brine and dried over anhydrous NaSO₄. The product was purified withP-TLC. 3 mg of the title compound was obtained. Yield: 22.2%. ¹H NMR(CDCl₃, 300 MHz) δ=6.73-6.61 (m, 4H), 5.95 (d, J=11.0 Hz 2H), 4.11 (d,J=15.3 Hz, 1H), 3.89 (s, 3H), 3.87 (s, 3H), 3.66-3.52 (m, 2H), 3.31-3.06(m, 3H), 2.86-2.60 (m, 3H). MS: m/z (APCI-ESI): 340.1 (M⁺+1).

50. Chiral Separation of Compound 1.

Compound 1 was subjected to chiral HPLC using the following HPLC system:

Column CHIRALCEL OJ-H Column size 0.46 cm I.D. × 15 cm L Injection 10 μlMobile phase Hexane/IPA/DEA = 95/5/0.1 (v/v/v) Flow rate 1.0 mL/min.Wave length UV 220 nm Temperature 35° C. Sample solution x mg/mL inMobile phase HPLC equipment Shimadzu LC 20 with UV detector SPD-20ASolvent Brand Tedia, HPLC gradeThe (R,S) and (S,R) enantiomers of compound 1 were separated andassigned as shown in the scheme. Under these conditions, the13S,14R-enantiomer (compound 31) exhibited a retention time of about 7minutes and the 13R14S-enantiomer (compound 32) exhibited a retentiontime of about 13 minutes.

51. Preparation of Compounds 23-26, 29.

To a solution of compound 34 (50 mg, 0.154 mmol) in acetone was addedR—Br (0.308 mmol), and K₂CO₃ (63.7 mg, 0.462 mmol), then the mixture washeated for 3 hours at about 70° C. in small ampoule. Then it waspurified by p-TLC to afford the product.

Compound 23: ¹H NMR (CDCl₃): δ 7.50-7.46 (m, 2H), 7.42-7.33 (m, 3H),6.90-6.80 (m, 2H), 6.72 (s, 1H), 6.58 (s, 1H), 5.92-5.91 (m, 2H), 5.03(dd, 2H, J₁=11.1 Hz, J₂=31.2 Hz), 4.20 (d, 1H, J=15.9 Hz), 3.87 (s, 3H),3.50-3.40 (m, 2H), 3.30-3.09 (m, 3H), 2.90-2.75 (m, 1H), 2.65-2.55 (m,2H). MS: m/z=416.0 (M⁺+1).

Compound 24: ¹H NMR (CDCl₃, 300 MHz): δ 7.43-7.35 (m, 3H), 7.19-7.16 (m,1H), 6.91-6.80 (m, 2H), 6.72 (s, 1H), 6.58 (s, 1H), 5.92-5.91 (m, 2H),5.03 (dd, 2H, J₁=11.1 Hz, J₂=31.2 Hz), 4.20 (d, 1H, J=15.9 Hz), 3.86 (s,3H), 3.52-3.47 (m, 2H), 3.30-3.05 (m, 3H), 2.90-2.75 (m, 1H), 2.65-2.55(m, 2H). MS: m/z=500.0 (M⁺+1).

Compound 25: ¹H NMR (CDCl₃): δ 7.50 (s, 1H), 7.35-7.29 (m, 3H),6.91-6.80 (m, 2H), 6.72 (s, 1H), 6.58 (s, 1H), 5.92-5.91 (m, 2H), 5.00(dd, 2H, J₁=11.1 Hz, J₂=31.2 Hz), 4.20 (d, 1H, J=15.9 Hz), 3.86 (s, 3H),3.55-3.40 (m, 2H), 3.30-3.05 (m, 3H), 2.90-2.75 (m, 1H), 2.65-2.55 (m,2H). MS: m/z=450.0 (M⁺+1).

Compound 26: ¹H NMR (CDCl₃, 300 MHz): δ 8.16-8.11 (m, 2H), 7.76-7.71 (m,2H), 7.75-7.46 (m, 2H), 7.93-7.80 (m, 2H), 6.73 (s, 1H), 6.58 (s, 1H),5.92-5.91 (m, 2H), 5.42-5.41 (m, 2H), 4.96 (s, 1H), 2.23 (d, 1H, J=15.9Hz), 3.80 (s, 3H), 3.60-3.50 (m, 2H), 3.30-3.05 (m, 3H), 2.90-2.75 (m,1H), 2.65-2.55 (m, 2H). MS: m/z=461.0 (M⁺+1).

Compound 29: ¹H NMR (CDCl₃, 300 MHz): δ 6.85-6.76 (m, 2H), 6.72 (s, 1H),6.58 (s, 1H), 5.59 (m, 1H), 5.92 (s, 2H), 4.27-4.26 (m, 2H), 4.22-4.03(m, 2H), 3.83 (s, 3H), 3.56-3.49 (m, 2H), 3.30-3.05 (m, 3H), 2.90-2.65(m, 3H), 1.38 (t, 3H, J=6.9 Hz). MS: m/z=354.1 (M⁺+1).

52. Preparation of Compounds 27 and 28.

To a solution of compound 34 (50 mg, 0.154 mmol) in CH₂Cl₂ was addedsulfonyl chloride (0.185 mmol), and Na₂CO₃ (20 mg), then the mixture wasstirred overnight at room temperature in small ampoule. Then it waspurified by p-TLC to afford the product.

Compound 27: ¹H NMR (CD₃OH, 300 MHz): δ 7.03 (d, J=8.4 Hz, 1H), 6.83 (d,J=8.7 Hz, 1H), 6.71 (s, 1H), 6.59 (s, 1H), 5.91 (dd, J=1.5, 2.1 Hz, 2H),4.27 (d, J=16.2 Hz, 1H), 4.00-3.89 (m, 1H), 3.87 (s, 3H), 3.83-3.60 (m,2H), 3.27-3.16 (m, 3H), 2.85-2.64 (m, 3H), 2.32-2.10 (m, 4H), 2.95-2.65(m, 4H); MS: m/z=326.1 (M⁺+1).

Compound 28: ¹H NMR (CDCl₃, 300 MHz): δ 7.55-7.7.51 (m, 5H), 7.06 (d,J=8.4 Hz, 1H), 6.87 (d, J=8.4 Hz, 1H), 6.70 (s, 1H), 6.57 (s, 1H), 5.08(dd, J=1.2, 2.1 Hz, 2H), 4.75 (q, J=16.5 Hz, 2H), 4.19 (d, J=15.9 Hz,3H), 3.64-3.54 (m, 2H), 3.26-3.3.02 (m, 3H), 2.86-2.77 (m, 1H),2.65-2.55 (m, 2H); MS: m/z=480.0 (M⁺+1).

53. Preparation of Compound 30.

To a stirred solution of NaOH (4 g, 100 mmol) in water (20 mL) was addedberberine chloride (2.0 g, 5.4 mmol) at room temperature. Then acetone(1.6 mL) was added slowly at same temperature and stirred for 1 hour.TLC analysis indicated the completion of the reaction. The reactionsolution was filtered, sufficiently washed with 80% methanol and driedto get 1.7 g of compound 30. ¹H NMR (CDCl₃): δ 7.13 (s, 1H); 6.77-6.75(m, 2H); 6.57 (s, 1H); 5.94-5.93 (m, 2H); 5.89 (s, 1H); 5.34-5.30 (m,1H); 3.89 (s, 3H); 3.84 (s, 3H); 3.36-3.30 (m, 2H); 3.11-3.04 (m, 1H);2.84-2.76 (m, 2H); 2.44-2.38 (m, 1H); 2.04 (s, 3H). MS: m/z (APCI-ESI)336.0 (M+H)⁺.

54. Preparation of Compounds 33 and 37.

To a solution of 1.0 g (2.7 mmol) of berberine chloride in 50 mL of MeOHwas added KCN (175 mg, dissolved in the minimum amount of water). Theyellow precipitate that immediately appeared was filtered and washedwith methanol. The solid was dried under vacuum. 810 mg of compound 33was obtained. Yield: 82.9%

¹H NMR (CDCl₃, 300 MHz): δ 7.17 (s, 1H), 6.86 (dd, J=8.7, 15.3 Hz, 2H),6.60 (s, 1H), 6.15 (s, 1H), 5.97 (dd, J=1.4, 5.1 Hz, 2H), 5.75 (s, 1H),3.97 (s, 1H), 3.87 (s, 1H), 3.49-3.38 (m, 1H), 3.30-3.24 (m, 1H),3.05-2.80 (m, 2H); MS: m/z=336.0 (M⁺−CN).

200 mg of compound 33 was dissolved in 1.0 mL of conc. HCl at 0° C. for30 minutes, then the mixture was dissolved in 15 mL of menthol. NaBH₄was added in portions until the yellow solution turned to colorless. Thereaction was quenched with water, extracted with DCM. The organic phasewas washed with water, brine and dried over anhydrous NaSO₄. The solventwas removed in vacuo, the residue was purified by column chromatographyto give compound 37. Yield: 78 mg (37.0%).

¹H NMR (CDCl₃, 300 MHz): δ 7.06 (s, 1H), 6.97 (s, 1H), 6.95-6.85 (m,3H), 6.65 (s, 1H), 5.94 (q, J=0.9 Hz, 2H), 4.01 (s, 1H), 3.77 (s, 3H),3.68 (s, 3H), 3.43-3.19 (m, 3H), 2.89-2.56 (m, 3H), 2.46-2.38 (m, 1H);MS: m/z=383.0 (M⁺+1)

55. Preparation of Compound 35.

A solution of 200 mg (0.59 mmol) of A (tetrahydroberberine) in 3 mL ofCHCl₃ was stirred with 110 mg (0.64 mmol) of m-CPBA at room temperatureovernight. The reaction mixture was diluted with 10 mL of DCM, washedwith 10% aqueous Na₂CO₃, dried and the solvent was evaporated. Theresidue was purified by column chromatography to give 185 mg of compound35. Yield: 85.2%.

¹H NMR (CDCl₃, 300 MHz): 6.98 (d, J=8.4 Hz, 1H), 6.87 (d, J=8.7 Hz, 1H),6.71 (s, 1H), 6.65 (s, 1H), 5.92 (dd, J=1.5, 4.8 Hz, 2H), 4.70-4.43 (m,3H), 3.93-3.89 (m, 1H), 3.86 (s, 3H), 3.85 (s, 3H), 3.83-3.80 (m, 1H),3.66-3.52 (m, 2H), 3.23 (dd, J=3.9, 16.2 Hz, 1H), 2.68 (dd, J=3.6, 16.5Hz, 1H); MS: m/z=356.0 (M⁺+1).

56. Preparation of Compounds 37, 38 and 42.

To a solution of A (50 mg, 0.15 mmol) in CH₂Cl₂ was added chlorosulfone(0.20 mmol), and two drops of Et₃N, then the mixture was stirred for 3hours at room temperature in a small ampoule. Then it was purified byp-TLC to afford the title compounds.

Compound 37: ¹H NMR (CDCl₃, 300 MHz): δ 8.09-7.44 (m, 9H), 7.02 (d,J=8.4 Hz, 1H), 6.72 (d, J=8.4 Hz, 1H), 6.71 (s, 1H), 6.59 (s, 1H), 5.92(s, 2H), 4.25 (d, J=15.9 Hz, 1H), 3.68-3.56 (m, 2H), 3.45 (s, 3H),3.28-3.08 (m, 3H), 2.87-2.60 (m, 3H); MS: m/z=542.1 (M⁺+1).

Compound 38: ¹H NMR (CDCl₃, 300 MHz): δ 7.93-7.55 (m, 4H), 7.01 (d,J=8.4 Hz, 1H), 6.72-6.69 (m, 3H), 5.91 (s, 2H), 4.19 (d, J=19.5 Hz, 1H),3.61-3.54 (m, 2H), 3.43 (s, 3H), 3.24 (dd, J=3.9, 15.9 Hz, 1H),3.12-3.02 (m, 2H), 2.86-2.51 (m, 3H); MS: m/z=522.1 (M⁺+1).

Compound 42: ¹H NMR (CDCl₃, 300 MHz): δ 7.93 (d, J=2.1 Hz, 1H),7.91-7.90 (d, J=2.4 Hz, 1H), 7.02-6.99 (m, 3H), 6.72 (d, J=9.6 Hz, 2H),6.58 (s, 1H), 5.91 (s, 2H), 4.23 (d, J=17.1 Hz, 1H), 3.90 (s, 3H),3.64-3.59 (m, 2H), 3.50 (s, 3H), 3.27-3.21 (m, 1H), 3.15-3.10 (m, 2H),2.85-2.59 (m, 3H); MS: m/z=490.0 (M⁺+1).

57. Preparation of Compound 39.

To a solution of A (50 mg, 0.15 mmol) in acetone was added2-bromopropane (38 mg, 0.31 mmol), and K₂CO₃ (63.7 mg, 0.46 mmol), thenthe mixture was heated for 3 hours at about 70° C. in a small ampoule.Then it was purified by p-TLC to afford the title compound 39. ¹H NMR(CDCl₃, 300 MHz): δ 6.79 (dd, J=8.1, 19.8 Hz, 2H), 6.72 (s, 1H), 6.58(s, 1H), 5.91 (s, 2H), 4.61-4.53 (m, 1H), 4.24 (d, J=15.9 Hz, 1H), 3.81(s, 3H), 3.53-3.50 (m, 2H), 3.24-3.10 (m, 3H), 2.86-2.80 (m, H),2.67-2.57 (m, 2H), 1.30 (d, J=2.7 Hz, 3H), 1.28 (d, J=2.4 Hz, 3H); MS:m/z=368.1 (M⁺+1).

58. Preparation of Compounds 41, 44, 45 and 46.

To a stirred solution of A (100 mg, 0.31 mmol) in DCM (1.5 mL) andpyridine (1.5 mL) was added B (33 μL, 0.34 mmol) at room temperature.Then the reaction mixture was heated to 40° C. and stirred for 8 hours.TLC analysis indicated the completion of the reaction. The solvent wasremoved in reduce pressure. The residue was extracted with EtOAc andwater, dried and concentrated to get crude product. The crude materialsufficiently washed with Et₂O and dried to get 35 mg of compound 41. ¹HNMR (CDCl₃): δ 6.97 (d, J=8.7 Hz, 1H); 6.81 d, J=8.7 Hz, 1H); 6.72 (s,1H); 6.58 (s, 1H); 5.91 (s, 2H); 5.01 (d, J=8.1 Hz, 1H); 4.07 (d, J=15.6Hz, 1H); 3.90-3.86 (m, 1H); 3.82 (s, 3H); 3.60-3.48 (m, 2H); 3.25-3.04(m, 3H); 2.86-2.77 (m, 1H); 2.66-2.55 (m, 2H); 1.24-1.12 (m, 6H). MS:m/z (APCI-ESI) 411.1 (M+H)⁺.

The compounds 44, 45 and 46 were prepared analogously using theappropriate isocyanate.

Compound 44: ¹H NMR (CDCl₃): δ 7.46 (dd, J=4.8 Hz, 2H), 7.16 (dd, J=8.4Hz, 2H), 7.04 (dd, J=8.4 Hz, 1H), 6.85 (dd, J=8.4 Hz, 1H), 6.73 (s, 1H),6.58 (m, 1H), 5.92 (s, 2H), 4.12-4.17 (m, 1H), 3.82 (s, 3H), 3.55-3.56(m, 2H), 3.14-3.25 (m, 3H), 2.98-2.75 (m, 1H), 2.61-2.61 (m, 2H). MS:m/z=529.0 (M⁺+1).

Compound 45: ¹H NMR (CDCl₃): δ 7.48 (m, 1H), 7.03 (dd, J=8.1 Hz, 1H),6.85 (dd, J=8.4 Hz, 1H), 6.75 (s, 1H), 6.67 (s, 2H), 6.59 (s, 1H), 6.19(s, 1H), 5.92 (s, 2H), 4.16-4.21 (m, 1H), 3.83 (s, 3H), 3.70 (s, 6H),3.48-3.64 (m, 2H), 3.13-3.31 (m, 3H), 2.83-2.92 (m, 1H), 2.57-2.83 (m,2H). MS: m/z=505.1 (M⁺+1).

Compound 46: ¹H NMR (CDCl₃): δ 7.4 (m, 1H), 7.16-7.23 (m, 2H), 7.03 (dd,1H, J=8.4 Hz), 6.83-6.90 (m, 2H), 6.75 (s, 2H), 6.58 (s, 1H), 6.19 (s,1H), 5.92 (s, 2H), 4.14-4.19 (m, 1H), 3.83 (s, 3H), 3.49-3.63 (m, 2H),3.08-3.31 (m, 3H), 2.81-2.90 (m, 1H), 2.57-2.67 (m, 2H), 2.29 (s, 3H).MS: m/z=459.1 (M⁺+1).

59. Preparation of Compound 43.

100 mg of A (0.31 mmol) was dissolved in 5.0 mL of DCM, and the solutionturned yellow. Then 0.5 mL of chloro-N,N-dimethylamide, 59 mg (0.307mmol) of EDCI and 1.0 mL of Et₃N were added and kept stirring overnightat reflux point. Cooled down and washed with water three times. Purifiedby silica gel chromatography to afford 45.0 mg of compound 43. ¹H NMR(CDCl₃): δ 6.97 (dd, J=8.4 Hz, 1H), 6.81 (dd, J=8.4 Hz, 1H), 6.71 (m,1H), 6.58 (s, 1H), 4.04-4.09 (m, 1H), 3.80 (m, 3H), 3.55-3.56 (m, 2H),3.02-3.25 (m, 8H), 2.61-2.79 (m, 4H). MS: m/z=397.1 (M⁺+1).60. Preparation of Compounds 49-53 and 56-60.

To a solution of A (50 mg, 0.15 mmol) in CH₂Cl₂ was added chlorosulfone(0.20 mmol), and two drops of Et₃N; then the mixture was stirred for 3hours at room temperature in a small ampoule. The reaction mixture waspurified by prep-TLC to afford the title compounds.

Compound 49: ¹H NMR (CDCl₃, 300 MHz): δ 8.00 (dd, J=5.7, 9.0 Hz, 1H),7.38 (dd, J=2.4, 8.4 Hz, 1H), 7.13-7.07 (m, 1H), 7.05 (d, J=8.4 Hz, 1H),6.71-6.68 (m, 2H), 6.59 (s, 1H), 5.91 (s, 2H), 4.26 (d, J=15.6 Hz, 1H),3.65-3.56 (m, 2H), 3.39 (s, 3H), 3.26-3.09 (m, 3H), 2.69-2.61 (m, 3H);MS: m/z=517.9 (M⁺+1).

Compound 50: ¹H NMR (CDCl₃, 300 MHz): δ 7.89-7.86 (dd, J=1.2, 7.6 Hz,1H), 7.55-7.50 (m, 1H), 7.41 (d, J=7.5 Hz, 1H), 7.32-7.27 (m, 1H), 6.99(d, J=8.4 Hz, 1H), 6.70-6.66 (m, 2H), 6.58 (s, 1H), 5.91 (s, 2H), 4.18(d, J=15.9 Hz, 1H), 3.59-3.54 (m, 2H), 3.32 (s, 3H), 3.26-3.07 (m, 3H),2.82 (s, 3H), 2.66-2.56 (m, 3H); MS: m/z=480.0 (M⁺+1).

Compound 51: ¹H NMR (CDCl₃, 300 MHz): δ 7.88-7.82 (m, 1H), 7.71-7.63 (m,1H), 7.34-7.28 (m, 2H), 7.01 (d, J=8.7 Hz, 1H), 6.68 (s, 1H), 6.68 (d,J=8.1 Hz, 1H), 6.59 (s, 1H), 5.91 (s, 2H), 4.27 (d, J=15.9 Hz, 1H),3.68-3.56 (m, 2H), 3.35 (s, 3H), 3.26-3.08 (m, 3H), 2.86-2.58 (m, 3H);MS: m/z=484.0 (M⁺+1).

Compound 52: ¹H NMR (CDCl₃, 300 MHz): δ 7.82-7.78 (m, 1H), 7.74-7.70 (m,1H), 7.59-7.52 (m, 1H), 7.42-7.36 (m, 1H), 7.03 (d, J=8.1 Hz, 1H),6.74-6.70 (m, 2H), 6.59 (s, 1H), 5.91 (s, 2H), 4.24 (d, J=15.9 Hz, 1H),3.67-3.57 (m, 2H), 3.45 (s, 3H), 3.28-3.09 (m, 3H), 2.86-2.59 (m, 3H);MS: m/z=484.0 (M⁺+1).

Compound 53: ¹H NMR (CDCl₃, 300 MHz): δ 7.91-7.77 (m, 2H), 7.40-7.32 (m,1H), 7.05 (d, J=8.4 Hz, 1H), 6.75-6.71 (m, 2H), 6.59 (s, 1H), 5.91 (s,2H), 4.25 (d, J=15.6 Hz, 1H), 3.70-3.58 (m, 2H), 3.50 (s, 3H), 3.27-3.05(m, 3H), 2.88-2.61 (m, 3H); MS: m/z 502.0 (M⁺+1).

Compound 56: ¹H NMR (CDCl₃, 300 MHz): δ 7.78-7.73 (m, 2H), 7.16-7.13 (m,1H), 7.04 (d, J=8.4 Hz, 1H), 6.77-6.70 (m, 2H), 6.59 (s, 1H), 5.91 (s,2H), 4.22 (d, J=15.9 Hz, 1H), 3.63-3.55 (m, 2H), 3.59 (s, 3H), 3.27-3.04(m, 3H), 2.86-2.60 (m, 3H); MS: m/z=472.0 (M⁺+1).

Compound 57: ¹H NMR (CDCl₃, 300 MHz): δ 7.89-7.85 (m, 2H), 7.36-7.33 (m,2H), 7.01 (d, J=8.7 Hz, 1H), 6.71 (d, J=7.2 Hz, 1H), 6.70 (s, 1H), 6.58(s, 1H), 5.91 (s, 2H), 4.20 (d, J=15.9 Hz, 1H), 3.62-3.54 (m, 2H), 3.45(s, 3H), 3.28-3.02 (m, 3H), 2.85-2.53 (m, 3H), 2.47 (s, 3H); MS:m/z=480.0 (M⁺+1)

Compound 58: ¹H NMR (CDCl₃, 300 MHz): δ 8.04-7.99 (m, 2H), 7.27-7.21 (m,2H), 7.02 (d, J=8.4 Hz, 1H), 6.71 (d, J=7.5 Hz, 1H), 6.70 (s, 1H), 6.59(s, 1H), 5.91 (s, 2H), 4.24 (d, J=15.9 Hz, 1H), 3.67-3.56 (m, 2H), 3.46(s, 3H), 3.28-3.03 (m, 3H), 2.86-2.59 (m, 3H); MS: m/z=484.0 (M⁺+1).

Compound 59: ¹H NMR (CDCl₃, 300 MHz): δ 7.90-7.82 (m, 1H), 7.08-6.97 (m,3H), 6.71-6.68 (m, 2H), 6.59 (s, 1H), 5.91 (s, 2H), 4.27 (d, J=15.9 Hz,1H), 3.69-3.55 (m, 2H), 3.40 (s, 3H), 3.26-3.05 (m, 3H), 2.85-2.59 (m,3H); MS: m/z=502.0 (M⁺+1).

Compound 60: ¹H NMR (CDCl₃, 300 MHz): δ 7.82-7.76 (m, 2H), 7.49-7.41 (m,2H), 7.01 (d, J=8.7 Hz, 1H), 6.72 (d, J=9.0 Hz, 1H), 6.70 (s, 1H), 6.58(s, 1H), 5.91 (s, 2H), 4.20 (d, J=15.9 Hz, 1H), 3.61-3.54 (m, 2H), 3.46(s, 3H), 3.27-3.04 (m, 3H), 2.85-2.55 (m, 3H), 2.45 (s, 3H); MS:m/z=480.0 (M⁺+1)

61. Preparation of Compound 47.

To a solution of 65.0 mg (0.2 mmol) of A in 2 mL of DCM and 1 mL ofpyridine was added 0.22 mmol of RNCO. The mixture was stirred at 40° C.for 6 hours. Then it was purified by p-TLC to afford the title compound.

Compound 47: ¹H NMR (CDCl₃): δ 8.12 (br, 1H), 7.70-7.58 (m, 2H),7.41-7.28 (m, 2H), 7.04 (d, J=8.7 Hz, 1H), 6.86 (d, J=8.4 Hz, 1H), 6.76(s, 1H), 6.58 (m, 1H), 5.94 (s, 2H), 4.26 (d, J=15.9 Hz, 1H), 3.85 (s,3H), 3.71-3.3.57 (m, 2H), 3.36-3.14 (m, 3H), 2.96-2.69 (m, 3H). MS:m/z=513.0 (M⁺+1).

62. Preparation of Compound 48.

To a solution of A (50 mg, 0.15 mmol) in CH₂Cl₂ was added ethylchloroformate (0.20 mmol), and two drops of Et₃N, then the mixture wasstirred for 3 hours at room temperature in a small ampoule. Then it waspurified by p-TLC to afford the title compound.

Compound 48: ¹H NMR (CDCl₃, 300 MHz): δ 7.01 (d, J=8.1 Hz, 1H), 6.83 (d,J=8.4 Hz, 1H), 6.58 (s, 1H), 5.91 (s, 2H), 4.36-4.29 (m, 2H), 4.12 (d,J=15.9 Hz, 1H), 3.83 (s, 3H), 3.57-3.48 (m, 2H), 3.26-3.11 (m, 3H),3.68-2.62 (m, 3H), 1.41-1.37 (m, 3H); MS: m/z=398.1 (M⁺+1).

63. Preparation of Compound 54.

To a stirred solution of A (50 mg, 0.15 mmol) in DCM (2 mL) and Et₃N (3drops) was added B (40 μL) at room temperature. Then the reactionmixture was heated to 40° C. and stirred for 3 hours. When TLC analysisindicated the completion of the reaction, the solvent was removed inreduced pressure. The residue was dissovled with EtOAc, This solutionwas washed with water, dried and concentrated to get crude product. Thecrude material was further purified by p-TLC to get 20 mg of compound54.

Compound 54: ¹H NMR (CDCl₃, 300 MHz): δ 8.26-8.23 (m, 2H), 7.68-7.63 (m,1H), 7.56-7.50 (m, 2H), 7.06 (d, J=9.3 Hz, 1H), 6.88 (d, J=8.4 Hz, 1H),6.74 (s, 1H), 6.57 (s, 1H), 5.92 (s, 2H), 4.08 (d, J=14.1 Hz, 1H), 3.79(s, 3H), 3.61-3.47 (m, 2H), 3.28-3.24 (m, 1H), 3.09-3.06 (m, 2H),2.91-2.81 (m, 1H), 2.64-2.56 (m, 2H); MS: m/z=430.0 (M++1).

64. Preparation of Compound 55.

To a stirred solution of A (50 mg, 0.15 mmol) and Et₃N (3 drops) in DCM(2 mL) was added B (40 μL) at room temperature. Then the reactionmixture was heated to 40° C. and stirred for 3 hours. When TLC analysisindicated the completion of the reaction, the solvent was removed inreduced pressure. The residue was dissolved with EtOAc. This solutionwas washed with water, dried and concentrated to get crude product. Thecrude material was further purified by p-TLC to get 26 mg of compound55.

Compound 55: ¹H NMR (CDCl₃, 300 MHz) δ 7.46-7.39 (m, 5H), 7.01 (d, J=8.4Hz, 1H), 6.83 (d, J=8.4 Hz, 1H), 6.69 (s, 1H), 6.58 (s, 1H), 5.91 (s,2H), 5.29 (s, 2H), 4.18 (d, J=12.6 Hz, 2H), 3.81-3.53 (m, 2H), 3.76 (s,3H), 3.28-3.18 (m, 3H), 2.94-2.68 (m, 3H); MS: m/z=460.1 (M⁺+1).

65. Preparation of Compound 61.

To a stirred solution of A (50 mg, 0.15 mmol) and pyridine (0.5 mL) inDCM (2 mL) was added B (20 mg, 0.17 mmol) at room temperature. Then thereaction mixture was heated to 40° C. and stirred for 6 hours. When TLCanalysis indicated the completion of the reaction, the solvent wasremoved in reduced pressure. The residue was dissolved with EtOAc. Thissolution was washed with water, dried and concentrated to get crudeproduct. The crude material was further purified by p-TLC to get 50 mgof compound 61. ¹H NMR (CDCl₃, 300 MHz) δ 7.45 (d, J=7.8 Hz, 2H), 7.30(t, J=7.5 Hz, 2H), 7.10-7.01 (m, 2H), 6.86 (d, J=8.4 Hz, 1H), 6.70 (s,1H), 6.58 (s, 1H), 5.93-5.92 (m, 2H), 4.32 (d, J=16.5 Hz, 1H), 3.83 (s,3H), 3.72 (br, 2H), 3.30-3.23 (m, 2H), 2.95-2.70 (m, 4H); MS: m/z=445.0(M⁺+1).

66. Preparation of Compound 62.

To a stirred solution of A (50 mg, 0.15 mmol) in DCM (2 mL) was addedbromopropane (21 mg, 0.17 mmol), Cs₂CO₃ (65 mg, 0.20 mmol) and KI (25mg, 0.15 mmol) at room temperature. Then the reaction mixture was heatedto 40° C. and stirred for 6 hours. When TLC analysis indicated thecompletion of the reaction, the solvent was removed in reduced pressure.The residue was dissolved with EtOAc. This solution was washed withwater, dried and concentrated to get crude product. The crude materialwas further purified by p-TLC to get 20 mg of compound 62. ¹H NMR(CDCl₃, 300 MHz) δ 6.81 (dd, J=8.4 Hz, 21 Hz, 2H), 6.72 (s, 1H), 6.59(s, 1H), 5.91 (t, J=1.5 Hz, 2H), 4.27 (d, J=15.6 Hz, 1H), 4.0-3.89 (m,2H), 3.82 (s, 3H), 3.59-3.55 (m, 2H), 3.25-3.19 (m, 3H), 2.88-2.80 (m,1H), 2.70-2.66 (m, 2H), 1.83-1.76 (m, 2H), 1.04 (t, J=7.2 Hz, 3H). MS:m/z=368.1 (M⁺+1).

67. Preparation of Compounds 63, 65 and 66.

To a stirred solution of A (50 mg, 0.15 mmol) in DCM (2 mL) was added B(0.17 mmol) at room temperature, Et₃N (0.1 mL) was added and stirred for4 hours. When TLC analysis indicated the completion of the reaction, thesolvent was removed in reduced pressure. The residue was furtherpurified by p-TLC to get title compounds.

Compound 63: ¹H NMR (CDCl₃, 300 MHz) δ 8.92 (t, J=2.1 Hz, 1H), 8.56-8.54(m, 1H), 8.39-8.34 (m, 1H), 7.80 (t, J=8.4 Hz, 1H), 7.38 (d, J=8.7 Hz,1H), 6.76-6.72 (m, 2H), 6.60 (s, 1H), 5.92 (s, 2H), 4.30 (d, J=15.6 Hz,1H), 3.72 (d, J=15.3 Hz, 1H), 3.62-3.59 (m, 1H), 3.50 (s, 3H), 3.28-3.05(m, 3H), 2.89-2.79 (m, 1H), 2.69-2.60 (m, 2H). MS: m/z=511.0 (M⁺+1).

Compound 65: ¹H NMR (CDCl₃, 300 MHz) δ 8.52-8.47 (m, 2H), 8.24-8.20 (m,2H), 7.11 (d, J=8.4 Hz, 1H), 6.94 (d, J=8.4 Hz, 1H), 6.92 (s, 1H), 6.68(s, 1H), 5.96 (s, 2H), 4.05 (d, J=15.9 Hz, 1H), 3.50-3.42 (m, 2H), 3.36(s, 3H), 3.06-2.84 (m, 2H), 2.67-2.42 (m, 4H); MS: m/z=511.0 (M⁺+1).

Compound 66: ¹H NMR (CDCl₃, 300 MHz) δ 9.26-9.24 (m, 1H), 8.41-8.12 (m,3H), 7.65-7.57 (m, 2H), 6.96 (d, J=8.4 Hz, 1H), 6.68 (s, 1H), 6.58 (d,J=9.9 Hz, 1H), 6.57 (s, 1H), 5.90 (s, 2H), 4.30 (d, J=15.9 Hz, 1H),3.64-3.52 (m, 2H), 3.23-3.00 (m, 3H), 3.02 (s, 3H), 2.83-2.49 (m, 3H);MS: m/z=517.0 (M⁺+1).

68. Preparation of Compound 64.

To 5.2 g of A was added phenylboric acid and toluene. The mixture washeated at reflux for 1 hour, and water was collected in a Dean-starktrap. The hot solution was poured over molecular sieves (3.7 g) in astainless steel bomb. Paraformaldehyde (6.4 g) was added. The bomb wassealed and heated on an oil-bath at 110° C. for 48 hours. The bomb wasopened and the hot solution filtered. The toluene was evaporated, andwater was added to the residue. After heating at reflux for 2 hours, themixture was cooled to room temperature and extracted with DCM. Thesolution was dried and the solvent was removed. The residue was washedwith ether to obtained 2.5 g of B.

To a solution of B (1.0 g, 5.2 mmol) in 20 mL of acetone was added K₂CO₃(810 mg, 5.8 mmol), followed by BnBr (0.7 mL, 5.8 mmol). The mixture wasstirred at room temperature overnight. Then the mixture was diluted withEtOAc, washed with water, brine, and dried over anhydrous Na₂SO₄. Thesolvent was removed under vacuum. The residue was purified by flashchromatography to provide 1.2 g of C.

To a solution of C (200 mg, 0.7 mmol) in 6 mL of MeOH was added3,4-dimethoxy phenethylamine (0.3 mL, 1.8 mmol) dropwise. The mixturewas stirred at room temperature for 3 hours. The reaction mixture wasdiluted with EtOAc, washed with H₂O, brine, dried over anhydrous Na₂SO₄.Then the solvent was removed under vacuum. The residue was purified withflash chromatography to give 310 mg of D.

Compound D (101 mg, 0.25 mmol) was suspended in 2 mL of toluene. Themixture was stirred under nitrogen. Then 0.15 mL of phosphoryl chloridewas added in one portion, and the mixture was heated under reflux for 2hours. The reaction mixture was cooled under nitrogen. Excess phosphorylchloride and toluene was evaporated under vacuum. The residue wasdissolved in methanol, then 100 mg of NaBH₄ was added in portions. Themixture was stirred at room temperature for 30 minutes. Then the solventwas removed, the residue was diluted with EtOAc, washed with H₂O, brine,dried over anhydrous Na₂SO₄. Then the solvent was removed under vacuum.The residue was purified with flash chromatography to give 42 mg of E(compound 64).

Compound 64: ¹H NMR (CDCl₃, 300 MHz), δ 7.50-7.30 (m, 5H), 6.86 (dd,J=8.4, 23.7 Hz, 2H), 6.73 (s, 1H), 6.61 (s, 1H), 5.04 (dd, J=11.4, 33.9Hz, 2H), 4.22 (d, J=15.9 Hz, 1H), 3.89 (s, 3H), 3.87 (s, 3H), 3.86 (s,3H), 3.55-3.42 (m, 2H), 3.30-3.11 (m, 3H), 2.89-2.58 (m, 3H); MS:m/z=432.1 (M⁺+1).

Example 9

The following compounds of Table 5 were synthesized according the aboveprocedures or slight modifications thereof and were characterized bymass spectroscopy. Each compound gave the expected MH⁺ peak in the massspectrum.

TABLE 5 Molecular Molecular MS: m/z Cmpd No. Structure Chemical NameFormula Weight [M⁺ + 1]* 1

9,10-dimethoxy-13-benzyl- 5,6,7,8,13,13a-hexahydro-2H-1,3-dioxolano[4,5- g]isoquinolino[3,2- a]isoquinoline C27H27NO4 429.51430.2 2

(13aR)-2,3,9,10-tetramethoxy- 5,6,7,8,13,13a- hexahydroisoquinolino[3,2-a]isoquinoline hydrochloride C21H25NO4 •HCl 355.43** 356.2 3

(13aR)-2,3,9,10-tetramethoxy- 5,6,7,8,13,13a- hexahydroisoquinolino[3,2-a]isoquinoline sulfate C21H25NO4 •H2SO4 355.43** 356.2 4

(13aR)-2,3,9,10-tetramethoxy- 5,6,7,8,13,13a- hexahydroisoquinolino[3,2-a]isoquinoline citrate C21H25NO4 •citrate 355.43** 356.2 5

(13aR)-2,3,9,10-tetramethoxy- 5,6,7,8,13,13a- hexahydroisoquinolino[3,2-a]isoquinoline maleate C21H25NO4 •maleate 355.43** 356.2 6

(13S,13aR)-2,3,9,10- tetramethoxy-13-methyl- 5,6,7,8,13,13a-hexahydroisoquinolino[2,1- b]isoquinoline hydrochloride C22H27NO4 •HCl369.45** 370.2 7

(13S,13aR)-2,3,9,10- tetramethoxy-13-methyl- 5,6,7,8,13,13a-hexahydroisoquinolino[2,1- b]isoquinoline sulfate C22H27NO4 •H2SO4369.45** 370.2 8

(13S,13aR)-2,3,9,10- tetramethoxy-13-methyl- 5,6,7,8,13,13a-hexahydroisoquinolino[2,1- b]isoquinoline citrate C22H27NO4 •citrate369.45** 370.2 9

(13S,13aR)-2,3,9,10- tetramethoxy-13-methyl- 5,6,7,8,13,13a-hexahydroisoquinolino[2,1- b]isoquinoline maleate C22H27NO4 •maleate369.45** 370.2 10

2,3,10,11-tetramethoxy- 5,6,7,8,13,13a- hexahydroisoquinolino[3,2-a]isoquinoline C21H25NO4 355.43 356.1 11

2,3-dimethoxy-5,6,7,8,14,14a- hexahydro-11H-1,3- dioxoleno[4,5-g]isoquinolino[2,1- b]isoquinoline C20H21NO4 339.39 340.1 12

(13S,13aR)-13-ethyl-2,3,10,11- tetramethoxy-5,6,7,8,13,13a-hexahydroisoquinolino[2,1- b]isoquinoline C23H29NO4 383.48 384.2 13

(13R,13aR)-13-ethyl-2,3,10,11- tetramethoxy-5,6,7,8,13,13a-hexahydroisoquinolino[2,1- b]isoquinoline C23H29NO4 383.48 384.2 14

(13R,13aR)-2,3-dimethoxy-8- oxo-5,6,7,13,13a-pentahydroisoquinolino[2,1- b]isoquinoline-13-carboxylic acid C20H19NO5353.37 354.1 15

(13S,13aR)-2,3-dimethoxy-8- oxo-5,6,7,13,13a-pentahydroisoquinolino[2,1- b]isoquinoline-13-carboxylic acid C20H19NO5353.37 354.1 16

2,3,11-trimethoxy- 5,6,7,8,13,13a- hexahydroisoquinolino[2,1-b]isoquinolin-10-ol C20H23NO4 341.40 342.1 17

2,3,10-trimethoxy- 5,6,7,8,13,13a- hexahydroisoquinolino[2,1-b]isoquinolin-11-ol C20H23NO4 341.40 342.1 18

8,9-dimethoxy-6,11,12,13,6a- pentahydro-2H-1,3- dioxoleno[4,5-h]isoquinolino[2,1- b]isoquinolin-14-one C20H17NO5 351.10 352.0 19

8,9-dimethoxy- 6,11,12,13,14,6a-hexahydro- 2H-1,3-dioxoleno[4,5-h]isoquinolino[2,1- b]isoquinoline C20H21NO4 339.14 340.1 20

13-ethyl-2,3,9,10- tetramethoxy-5,6,7,8,13,13a-hexahydroisoquinolino[2,1- b]isoquinoline C23H29NO4 383.49 402.1 [M⁺ +1 + H₂O] 21

12-bromo-2,3,9-trimethoxy- 5,6,7,8,13,13a- hexahydroisoquinolino[2,1-b]isoquinoline C20H22BrNO3 404.31 403.9, 405.9 22

2,3,9-trimethoxy- 5,6,7,8,13,13a- hexahydroisoquinolino[3,2-a]isoquinoline C20H23NO3 325.41 326.1 23

10-methoxy-9- (phenylmethoxy)- 5,6,7,8,13,13a-hexahydro-2H-1,3-dioxolano[4,5- g]isoquinolino[3,2- a]isoquinoline C26H25NO4 415.48416.0 24

10-methoxy-9-{[3- (trifluoromethoxy)phenyl]meth- oxy}-5,6,7,8,13,13a-hexahydro-2H-1,3- dioxolano[4,5- g]isoquinolino[3,2- a]isoquinolineC27H24F3NO5 499.48 500.0 25

9-[(3-chlorophenyl)methoxy]- 10-methoxy-5,6,7,8,13,13a-hexahydro-2H-1,3- dioxolano[4,5- g]isoquinolino[3,2- a]isoquinolineC26H24ClNO4 449.93 450.0, 452.0 26

10-methoxy-9-[(2- nitrophenyl)methoxy]- 5,6,7,8,13,13a-hexahydro-2H-1,3-dioxolano[4,5- g]isoquinolino[3,2- a]isquinoline C26H24N2O6 460.48461.0 27

10-methoxy-5,6,7,8,13,13a- hexahydro-2H-1,3- dioxolano[4,5-g]isoquinolino[3,2- a]isoquinolin-9-yl cyclopentanesulfonate C24H27NO6S457.54 458.1 28

10-methoxy-5,6,7,8,13,13a- hexahydro-2H-1,3- dioxolano[4,5-g]isoquinolino[3,2- a]isoquinolin-9-yl benzylsulfonate C26H25NO6S 479.54480.0 29

9-ethoxy-10-methoxy- 5,6,7,8,13,13a-hexahydro-2H- 1,3-dioxolano[4,5-g]isoquinolino[3,2- a]isoquinoline C21H23NO4 353.41 354.1 30

1-(9,10-dimethoxy-5,6,7,8- tetrahydro-2H-1,3- dioxolano[4,5-g]isoquinolino[3,2- a]isoquinolin-8-yl)acetone C23H23NO5 393.43 336.0[M⁺ + 1-acetone] 31

(13S,13aR)-9,10-dimethoxy- 13-benzyl-5,6,7,8,13,13a- hexahydro-2H-1,3-dioxolano[4,5- g]isoquinolino[3,2- a]isoquinoline C27H27NO4 429.51 430.132

(13aS,13R)-9,10-dimethoxy- 13-benzyl-5,6,7,8,13,13a- hexahydro-2H-1,3-dioxolano[4,5- g]isoquinolino[3,2- a]isoquinoline C27H27NO4 429.51 430.133

9,10-dimethoxy-5,6,7,8- tetrahydro-2H-1,3- dioxolano[4,5-g]isoquinolino[3,2- a]isoquinoline-8-carbonitrile C21H18N2O4 362.38336.0 [M⁺ − CN] 34

10-methoxy-5,6,7,8,13,13a- hexahydro-2H-1,3 dioxolano[4,5-g]isoquinolino[3,2- a]isoquinolin-9-ol C19H19NO4 325.13 326.0 35

9,10-dimethoxy- 5,6,7,8,13,13a-hexahydro-2H- 1,3-dioxolano[4,5-g]isoquinolino[3,2- a]isoquinoline-N-oxide C20H21NO5 355.38 356.0 36

9,10-dimethoxy- 5,6,7,8,13,13a-hexahydro-2H- 1,3-dioxolano[4,5-g]isoquinolino[3,2- a]isoquinoline-8-carboxamide C21H22N2O5 382.41 383.037

10-methoxy-5,6,7,8,13,13a- hexahydro-2H-1,3- dioxolano[4,5-g]isoquinolino[3,2- a]isoquinolin-9-yl-4- phenylbenzenesulfonateC31H27NO6S 541.61 542.0 38

10-methoxy-5,6,7,8,13,13a- hexahydro-2H-1,3- dioxolano[4,5-g]isoquinolino[3,2- a]isoquinolin-9-yl-4-(tert- butyl)benzenesulfonateC29H31NO6S 521.62 522.1 39

10-methoxy-9-(methylethoxy)- 5,6,7,8,13,13a-hexahydro-2H-1,3-dioxolano[4,5- g]isoquinolino[3,2- a]isoquinoline C22H25NO4 367.44368.1 41

(10-methoxy(5,6,7,8,13,13a- hexahydro-2H-1,3- dioxolano[4,5-g]isoquinolino[3,2- a]isoquinolin-9-yloxy))-N- (methylethyl)carboxamideC23H26N2O5 410.46 411.1 42

10-methoxy-5,6,7,8,13,13a- hexahydro-2H-1,3- dioxolano[4,5-g]isoquinolino[3,2- a]isoquinolin-9-yl 4- methoxybenzenesulfonateC26H25NO7S 495.54 496.0 43

(10-methoxy(5,6,7,8,13,13a- hexahydro-2H-1,3- dioxolano[4,5-g]isoquinolino[3,2- a]isoquinolin-9-yloxy))-N,N- dimethylcarboxamideC22H24N2O5 396.44 397.1 44

(10-methoxy(5,6,7,8,13,13a- hexahydro-2H-1,3- dioxolano[4,5-g]isoquinolino[3,2- a]isoquinolin-9-yloxy))-N-[4-(trifluoromethoxy)phenyl] carboxamide C27H23F3N2O6 528.41 529.1 45

N-(3,5-dimethoxyphenyl)(10- methoxy(5,6,7,8,13,13a- hexahydro-2H-1,3-dioxolano[4,5- g]isoquinolino[3,2- a]isoquinolin-9- yloxy))carboxamideC28H28N2O7 504.53 505.1 46

(10-methoxy(5,6,7,8,13,13a- hexahydro-2H-1,3- dioxolano[4,5-g]isoquinolino[3,2- a]isoquinolin-9-yloxy))-N-(3-methylphenyl)carboxamide C27H26N2O5 458.51 459.1 47

(10-methoxy(5,6,7,8,13,13a- hexahydro-2H-1,3- dioxolano[4,5-g]isoquinolino[3,2- a]isoquinolin-9-yloxy))-N-[3-(trifluoromethoxy)phenyl] carboxamide C27H23F3N2O5 512.48 513.0 48

10-methoxy-5,6,7,8,13,13a- hexahydro-2H-1,3- dioxoalano[4,5-g]isoquinolino[3,2- a]isoquinolin-9-yl ethoxyformate C22H23NO6 397.42398.1 49

10-methoxy-5,6,7,8,13,13a- hexahydro-2H-1,3- dioxolano[4,5-g]isoquinolino[3,2- a]isoquinolin-9-yl 2chloro-4- fluorobenzenesulfonateC25H21ClFNO6S 517.95 517.9, 519.9 50

10-methoxy-5,6,7,8,13,13a- hexahydro-2H-1,3- dioxolano[4,5-g]isoquinolino[3,2- a]isoquinolin-9-yl 2- methylbenzenesulfonateC26H25NO6S 479.54 480.0 51

10-methoxy-5,6,7,8,13,13a- hexahydro-2H-1,3- dioxolano[4,5-g]isoquinolino[3,2- a]isoquinolin-9-yl 2- fluorobenzenesulfonateC25H22FNO6S 483.51 484.0 52

10-methoxy-5,6,7,8,13,13a- hexahydro-2H-1,3- dioxolano[4,5-g]isoquinolino[3,2- a]isoquinolin-9-yl 3- fluorobenzenesulfonateC25H22FNO6S 483.51 484.0 53

10-methoxy-5,6,7,8,13,13a- hexahydro-2H-1,3- dioxolano[4,5-g]isoquinolino[3,2- a]isoquinolin-9-yl 3,4- difluorobenzenesulfonateC25H21F2NO6S 501.50 502.0 54

10-methoxy-5,6,7,8,13,13a- hexahydro-2H-1,3- dioxolano[4,5-g]isoquinolino[3,2- a]isoquinolin-9-yl benzoate C26H23NO5 429.46 430.055

10-methoxy-5,6,7,8,13,13a- hexahydro-2H-1,3- dioxolano[4,5-g]isoquinolino[3,2- a]isoquinolin-9-yl (phenylmethoxy)formate C27H25NO6459.49 460.1 56

10-methoxy-5,6,7,8,13,13a- hexahydro-2H-1,3- dioxolano[4,5-g]isoquinolino[3,2- a]isoquinolin-9-yl thiophene-2- sulfonateC23H21NO6S2 471.55 472.0 57

10-methoxy-5,6,7,8,13,13a- hexahydro-2H-1,3- dioxolano[4,5-g]isoquinolino[3,2- a]isoquinolin-9-yl 4- methylbenzenesulfonateC26H25NO6S 479.54 480.0 58

10-methoxy-5,6,7,8,13,13a- hexahydro-2H-1,3- dioxolano[4,5-g]isoquinolino[3,2- a]isoquinolin-9-yl 4- fluorobenzenesulfonateC25H22FNO6S 483.51 484.0 59

10-methoxy-5,6,7,8,13,13a- hexahydro-2H-1,3- dioxolano[4,5-g]isoquinolino[3,2- a]isoquinolin-9-yl 2,4- difluorobenzenesulfonateC25H21F2NO6S 501.50 502.0 60

10-methoxy-5,6,7,8,13,13a- hexahydro-2H-1,3- dioxolano[4,5-g]isoquinolino[3,2- a]isoquinolin-9-yl 3- methylbenzenesulfonateC26H25NO6S 479.54 480.0 61

(10-methoxy(5,6,7,8,13,13a- hexahydro-2H-1,3- dioxolano[4,5-g]isoquinolino[3,2- a]isoquinolin-9-yloxy))-N- benzamide C26H24N2O5444.48 445.0 62

10-methoxy-9-propoxy- 5,6,7,8,13,13a-hexahydro-2H- 1,3-dioxolano[4,5-g]isoquinolino[3,2- a]isoquinoline C22H25NO4 367.44 368.1 63

10-methoxy-5,6,7,8,13,13a- hexahydro-2H-1,3- dioxolano[4,5-g]isoquinolino[3,2- a]isoquinolin-9-yl 3- nitrobenzenesulfonateC25H22N2O8S 510.52 511.0 64

2,3,10-trimethoxy-9- (phenylmethoxy)- 5,6,7,8,13,13a-hexahydroisoquinolino[3,2- a]isoquinoline C27H29NO4 431.52 432.1 65

10-methoxy-5,6,7,8,13,13a- hexahydro-2H-1,3- dioxolano[4,5-g]isoquinolino[3,2- a]isoquinolin-9-yl 4- nitrobenzenesulfonateC25H22N2O8S 510.52 511.0 66

10-methoxy-5,6,7,8,13,13a-hexahydro-2H-1,3- dioxolano[4,5-g]isoquinolino[3,2- a]isoquinolin-9-yl quinoine-8- sulfonate C28H24N2O6S516.56 517.0 67

[3,9,10-trimethoxy-5,8,13,13a- tetrahydro-6H-isoquino[3,2-a]isoquinolin-3-yl]-5-(2-oxo- hexahydro-1H-thieno[3,4-d]imidazol-4-yl)pentanoic ester C30H37N3O6S 567.24 568.3 68

2-[3,9,10-trimethoxy- 5,8,13,13a-tetrahydro-6H-isoquino[3,2-a]isoquinolin-3- yl]-acetic acid C22H25NO6 399.17 400.2 69

9-benzenesulfonyloxy-10- methoxy-5,8,13,13a- tetrahydro-6H-[1,3]dioxolo[4,5- g]isoquino[3,2-a]isoquinoline C25H23NO6S 465.12 466.270

9-benzoxy-10-methoxy- 5,8,13,13a-tetrahydro-6H- [1,3]dioxolo[4,5-g]isoquino[3,2-a]isoquinoline C20H21NO6S 403.11 404.1 71

9-methanesulfonyloxy-10- methoxy-5,8,13,13a- tetrahydro-6H-[1,3]dioxolo[4,5- g]isoquino[3,2-a]isoquinoline C20H21NO6S 403.11 404.172

9-benzenesulfonyloxy-10- methoxy-5,8,13,13a- tetrahydro-6H-[1,3]dioxolo[4,5- g]isoquino[3,2-a]isoquinoline C25H24ClNO6S 466.13466.2 73

10-methoxy-5-8-13-13a- tetrahydro-6H- [1,3]dioxolo[4,5-g]isoquino[3,2-a]isoquinoline- 9-ol C19H19NO4 325.13 326.2 74

9-O-3-(1'-bromo-propyl)-10- methoxy-5,8,13,13a- tetrahydro-6H-[1,3]dioxolo[4,5- g]isoquino[3,2-a]isoquinoline C22H24BrNO4 445.09 446.275

9-hydroxy-10-methoxy-5,6- dihydro-[1,3]dioxolo[4,5- g]isoquino[3,2-a]isoquinolinylium C19H16ClNO4 357.08 323.1 76

9-O-3-(1'-bromo-propyl)-10- methoxy-5,6-dihydro- [1,3]dioxolo[4,5-g]isoquino[3,2- a]isoquinolinylium C22H21BrClNO4 479.03 443.9 77

5,8,13,13a-Tetrahydro-6H- isoquino[3,2-a]isoquinoline- 2,3,9,10-tetraol;hydrobromide C17H18BrNO4 379.04 298.3 78

13-methyl-5,8,13,13a- Tetrahydro-6H-isoquino[3,2-a]isoquinoline-2,3,9,10-tetraol; hydrobromide C18H20BrNO4 393.06 313.879

9-benzenesulfonyloxy-10- methoxy-5,6-dihydro- [1,3]dioxolo[3,2-a]isoquinolinylium C25H20ClNO6S 497.07 462.2 80

9-methanesulfonyloxy-10- methoxy-5,6-dihydro- [1,3]dioxolo[4,5-g]isoquino[3,2- a]isoquinolinylium C20H18ClNO6S 435.05 399.5 81

9-benzoxy-10-methoxy-5,6- dihydro-[1,3]dioxolo[4,5- g]isoquino[3,2-a]isoquinolinylium C26H20ClNO5 461.1 426.1 82

9-O-2-(1'-hydroxy-ethyl)-10- methoxy-5,8,13,13a- tetrahydro-6H-[1,3]dioxolo[4,5- g]isoquino[3,2-a]isoquinoline C21H23NO5 369.16 370.183

2-[10-methoxy-5,8,13,13a- tetrahydro-6H- [1,3]dioxolo[4,5-g]isoquino[3,2-a]isoquinoline- 9-yl]-acetic esate C23H25NO6 411.7 412.284

9-(4-chloro- benzenesulfonyloxy)-10- methoxy-5,8,13,13a- tetrahydro-6H-[1,3]dioxolo[4,5- g]isoquino[3,2-a]isoquinoline C25H22ClNO6S 499.09500.3 85

9-(4-trifluoromethyl- benzenesulfonyloxy)-10- methoxy-5,8,13,13a-tetrahydro-6H- [1,3]dioxolo[4,5- g]isoquino[3,2-a]isoquinolineC26H22F3NO6S 533.52 534.2 86

9-(3,4-dimethoxy- benzenesulfonyloxy)-10- methoxy-5,8,13,13a-tetrahydro-6H- [1,3]dioxolo[4,5- g]isoquino[3,2-a]isoquinolineC27H27NO8S 525.57 526.3 87

9-(4-methylsulfonyl- benzenesulfonyloxy)-10- methoxy-5,8,13,13a-tetrahydro-6H- [1,3]dioxolo[4,5- g]isoquino[3,2-a]isoquinolineC26H25NO8S2 543.61 544.3 88

9-O-2-(1′-morpholine-ethyl)- 10-methoxy-5,8,13,13a- tetrahydro-6H-[1,3]dioxolo[4,5- g]isoquino[3,2-a]isoquinoline C25H30N2O5 438.52 439.289

10-methoxy-5,6,7,8,13,13a- hexahydro-2H-1,3-dioxolano[4,5-g]isoquinolino[3,2-a] isoquinolin-9-yl 5- (dimethylamino)naphthalenesulfonate C31H30N2O6S 558.64 559.1 90

10-methoxy-5,6,7,8,13,13a- hexahydro-2H-1,3-dioxolano[4,5-g]isoquinolino[3,2-a] isoquinolin-9-yl (trifluoromethyl)sulfonateC20H18F3NO6S 457.42 458.0 91

2,3,10-trimethoxy- 5,6,7,8,13,13a- hexahydroisoquinolino[2,1-b]isoquinollin-9-yl benzenesulfonate C26H27NO6S 481.56 482.0 92

10-methoxy-5,6,7,8,13,13a- hexahydro-2H-1,3-dioxolano[4,5-g]isoquinolino[3,2-a] isoquinoline C19H19NO3 309.36 310.0 93

10-methoxy-5,6,7,8,13,13a- hexahydro-2H-1,3-dioxolano[4,5-g]isoquinolino[3,2-a] isoquinolin-9-yl 2-(acetylamino)-4-methyl-1,3- thiazole-5-sulfonate C25H25N3O7S2 543.61543.9 94

10-methoxy-9-(3-methyl-5- nitro(2-pyridyl)oxy)- 5,6,7,8,13,13a-hexahydro-2H-1,3-dioxolano [4,5-g]isoquinolino[3,2-a] isoquinolineC25H23N3O6 461.47 462.0 95

2,10-dimethoxy-3- (phenylmethoxy)- 5,6,7,8,13,13a- hexahydroisoquinolino[2,1- b]isoquinolin-9-yl benzenesulfonate C32H31NO6S 557.66 558.0 96

10-methoxy-9-(5-nitro(2- pyridyl)oxy)-5,6,7,8,13,13a-hexahydro-2H-1,3-dioxolano [4,5-g]isoquinolino[3,2- a]isoquinolineC24H21N3O6 447.40 448.0 97

10-methoxy-5,6,7,8,13,13a- hexahydro-2H-1,3-dioxolano[4,5-g]isoquinolino [3,2- a]isoquinolin-9-yl thiophene-2- sulfonatehydrochloride C23H22ClNO6S2 508.00 471.9 98

10-methoxy-5,6,7,8,13,13a- hexahydro-2H-1,3-dioxolano[4,5-g]isoquinolino [3,2- a]isoquinolin-9-yl 3- fluorobenzenesulfonatehydrochloride C25H23ClFNO6S2 519.96 484.0 99

10-methoxy-5,6,7,8,13,13a- hexahydro-2H-1,3-dioxolano[4,5-g]isoquinolino [3,2- a]isoquinolin-9-yl 3,4-difluorobenzenesulfonate hydrochloride C25H22ClF2NO6S 537.95 502.0 100

6-(10-methoxy (5,6,7,8,13,13a- hexahydro-2H-1,3- dioxolano[4,5-g]isoquinolino[3,2- a]isoquinolin-9-yloxy))-5- methyl-3-pyridylamineC25H25N3O4 431.48 432.1 101

10-methoxy-5,6,7,8,13,13a- hexahydro-2H-1,3-dioxolano[4,5-g]isoquinolino [3,2- a]isoquinolin-9-yl pyridine-3- sulfonateC24H22N2O6S 466.51 467.0 102

3-Hydroxy-2,10-dimethoxy- 5,6,7,8,13,13a- hexahydroisoquinolino[2,1-b]isoquinolin-9-yl benzenesulfonate C25H25NO6S 467.53 468.0 103

2,10-Dimethoxy-8-prop-2- enyl-3-prop-2-enyloxy- 5,6,7,8,13,13a-hexahydroisoquinolino[2,1- b]isoquinolin-9-yl benzenesulfonateC31H33NO6S 547.66 548.1 104

10-methoxy-5,6,7,8,13,13a- hexahydro-2H-1,3-dioxolano[4,5-g]isoquinolino [3,2- a]isoquinolin-9-yl 3- fluorobenzenesulfonatehydrochloride C25H23ClFNO6S 519.97 512.0 105

9-O-2-(1′-phenylsulfonyl- ethyl)-10-methoxy-5,8,13,13a- tetrahydro-6H-[1,3]dioxolo[4,5- g]isoquino[3,2-a]isoquinoline C27H27NO6S 493.57 494.3*unless a different ion is indicated **molecular weight of free base

Example 10 Pharmacokinetics of CRDL

The plasma concentration of CRDL was determined in male hamstersfollowing intravenous or oral administration. Hamsters were given CRDLat 2 mg/kg i.v. or 20 mg/kg p.o. Blood samples were collected at theindicated time points and the plasma was isolated. Concentrations ofCRDL in plasma samples were determined by HPLC analysis, and the resultsare presented in Table 6 and FIGS. 7-8. Calculated pharmacokineticparameters are presented in Table 7. CRDL had a relatively short halflife (1.4 hr) and high clearance (5.6 L/hr/kg liver blood flow). It wasalso absorbed very quickly (T_(max)˜0.25 hr) and had a good exposure(F˜30%).

TABLE 6 Plasma Concentration of CRDL in Male Hamsters FollowingIntravenous and Oral Administration Time (hr) Concentration in plasma(ng/mL) IV-2 mg/kg H1 H2 H3 Mean S.D. 0 BLQ BLQ BLQ NA NA    0.083621.33 520.38 641.16 594.29 64.77   0.25 255.56 350.21 318.96 308.2548.23   0.5 124.95 165.87 130.79 140.54 22.13 1 60.71 69.78 69.09 66.535.05 2 23.92 19.37 27.99 23.76 4.31 6 6.34 4.76 4.67 5.25 0.94 24  BLQBLQ BLQ NA NA PO-20 mg/kg H4 H5 H6 Mean S.D. 0 BLQ BLQ BLQ NA NA   0.251263.21 895.64 1308.39 1155.75 226.39   0.75 369.71 164.19 217.42 250.44106.67 1 254.54 146.76 199.83 200.37 53.89 2 98.97 62.78 81.27 81.0118.10 6 14.54 9.66 20.83 15.01 5.60 8 10.93 6.80 18.54 12.09 5.96 24 3.41 2.64 3.12 3.04 0.42

TABLE 7 Selected Pharmacokinetic Parameters of CRDL in HamstersFollowing Intravenous and Oral Administration AUC_((0-t)) AUC_((0-∞))MRT_((0-∞)) t_(1/2z) T_(max) Vz CLz C_(max) μg/L * hr μg/L * hr hr hr hrL/kg L/hr/kg μg/L F % IV (2 mg/kg) H1 335.91 351.09 1.17 1.66 0.08 13.645.70 621.33 H2 336.84 345.02 0.94 1.19 0.08 9.96 5.80 520.38 H3 364.46373.56 0.98 1.35 0.08 10.44 5.35 641.16 mean 345.74 356.55 1.03 1.400.08 11.35 5.62 594.29 SD 16.22 15.03 0.12 0.24 0.00 2.00 0.23 64.77 PO(20 mg/kg) H4 1188.19 1232.21 3.41 8.94 0.25 NA NA 1263.21 34.56 H5756.88 794.56 3.96 10.13 0.25 NA NA 895.64 22.28 H6 1154.55 1183.46 3.376.43 0.25 NA NA 1308.39 33.19 mean 1033.21 1070.08 3.58 8.50 0.25 NA NA1155.75 30.01 SD 239.89 239.85 0.33 1.89 0.00 NA NA 226.39 6.73 NA: Notapplicable

Example 11 Evaluation of Pharmacokinetics of New Compound 72 Enantiomersin Male Wister Rats Following Intravenous and Oral Administration

The objective of this study was to collect plasma samples from maleWister rats at various time points following intravenous and oraladministration of test compounds (72(+) and 72(−)). These samples wereused later for the determination of plasma compound levels by LC/MS/MSfor estimating pharmacokinetic parameters.

Male Wister rats (body weight: 100 to 200 g) were used in this study.Before the pharmacokinetic studies, animals were randomly assigned to 4groups (3 animals per timepoint). The treatment condition is shown inTable 8.

For intravenous administration, blood samples were collected viaretro-orbital puncture into heparinized tubes at pre-dose and 0.083,0.25, 0.5, 1, 2, 6, 8, and 24 hours (hr) post-dose.

For oral administration, blood samples were collected at following timepoints: 0, 0.167, 0.417, 0.75, 1, 2, 4, 6, 8, and 24 hr. After samplecollection, plasma samples were stored at −20° C. until bioanalysis.Concentrations of 72(+) and 72(−) in plasma samples were determinedusing a high performance liquid chromatography/mass spectrometry(HPLC/MS/MS) method.

Table 9 lists selected non compartmental pharmacokinetic parameters ofenantiomer 72(+) in male rats following intravenous and oraladministration. Table 10 lists selected non compartmentalpharmacokinetic parameters of enantiomer 72(−) in male rats followingintravenous and oral administration.

Collectively, these results indicate that both enantiomers of newcompound 72 are relatively stable with half-lives of systemicclearance >2.5 hours and bioavailability higher than 45%. Thebioavailability (F*) was calculated by applying the following formula:*F(%)=(Dose_(iv) ×AUC _(oral(0-t)))/(Dose_(oral) ×AUC _(iv(0-t)))×100%.

TABLE 8 Experimental Design Treatment Number of Dose Dose Dose AnimalsLevel Conc Volume Dosing Group Male Test Article (mg/kg) (mg/ml) (mL/kg)Vehicle Route 1 3 72(+) 1 0.5 2 1% DMSO in 0.1 M IV phosphate buffer, pH3.0 2 3 72(+) 5 0.5 10 1% DMSO in 0.1 M PO phosphate buffer, pH 3.0 3 372(−) 1 0.5 2 1% DMSO in 0.1 M IV phosphate buffer, pH 3.0 4 3 72(−) 50.5 10 1% DMSO in 0.1 M PO phosphate buffer, pH 3.0 buffer, pH 3.0)

Example 12 In Vivo Studies for (14R)-THP

To determine whether the induction of hepatic LDLR expression bycorydalis-derived compounds would result in total cholesterol (TC) andLDL-cholesterol (LDL-c) reductions in plasma, hypercholesterolemichamsters were used as an animal model.

In one experiment, 10 Golden Syrian male hamsters under a highcholesterol (HC) diet were divided into 2 groups. One group was treatedwith (14R)-THP at a daily dose of 12 mg/kg by intra peritoneal (i.p.)injection and the control group received equal amount of vehicle by i.p.FIG. 9A shows the serum lipid levels of control and (14R)-THP treatmentgroups after 34 days of drug treatment. (14R)-THP reduced plasma TC by31.6%, TG by 35.93%, and non-HDL-c by 35.95% compared to the untreatedcontrol group. No adverse effects were observed during the treatmentperiod. FIG. 9B shows that gains of body weight during the treatmentperiod were very similar in control and drug-treated groups.

Example 13 In Vivo Studies for (14R,13S)-CRDL

In another experiment, 18 Golden Syrian male hamsters under a highcholesterol (HC) diet were divided into two treatment groups of i.p. Onegroup was treated with (14R,13S)-CRDL at a daily dose of 10 mg/kg andthe control group received equal amount of vehicle. FIG. 10A shows theplasma lipid levels of different groups after 17 days of drug treatment.(14R,13S)-CRDL reduced plasma TC by 41.9%, TG by 46.0%, and non-HDL-c by50% compared to the untreated group.

The HC diet markedly damaged the liver function of the hamsters, whichwas evident by elevated alanine aminotransaminase (ALT) and aspartateaminotransaminase (AST) in blood. (14R,13S)-CRDL treatment amelioratedthe liver function under the HC diet and lowered the serum level of ALTby 60% and lowered the serum level of AST by 30% compared to control(FIG. 10B). FIG. 10C showed that the blood levels of urea nitrogen (BUN)and glucose were not changed, while the level of creatinine (CREA) wasreduced by 20% (p<0.001). Again, no adverse effects were observed duringthe drug treatment.

Example 14 Lipid Lowering Effects of (14R,13S)-CRDL Hydrochloride Saltin Wister Rats

Male Wister rats were used as another animal model to examine the invivo effects of (14R,13S)-CRDL in plasma lipid levels.

In one experiment, 18 Wister male rats under a high fat and highcholesterol diet were divided into 2 groups. One group was given(14R,13S)-CRDL hydrochloride salt orally at a daily dose of 75 mg/kg byan orogastric tube for 4 weeks, and the control group received dailytreatment of equal amount of vehicle (0.1 M sodium phosphate, pH 3.5).Serum lipid levels were measured weekly. CRDL treatment resulted in astrong time-dependent reduction of serum lipid levels.

FIG. 11A shows that CRDL treatment lowered TC to 33.8% compared to thecontrol group and to 33.0% of the pretreatment level.

FIG. 11B shows that the LDL-c level was reduced by CRDL to 25.6% ofcontrol, and to 22.4% of day 0 by CRDL treatment.

FIG. 11C shows that the TG level was decreased to 29% of the control andto 27% of the pretreatment level (day 0).

No any adverse effects were observed during the entire treatment. At theend of treatment, Rats were sacrificed and blood and sera were collectedto measure several critical parameters for liver function and kidneyfunction. FIG. 11D showed that the serum levels of AST and ALT werestatistically lower in CRDL-treated group than control group, indicatingthat liver function was not damaged and was improved instead withstatistical significance. Kidney function and blood glucose level werenot changed by the treatment (FIG. 11E).

Example 15 Body-weight-reducing Effect of (4R,13S)-CRDL HydrochlorideSalt in Wister Rats

The food intake and body weight gains were measured every week duringthe 4-week treatment period. FIG. 12A shows the food intake during thetreatment. In the first week, the amount of food consumed was decreasedin CRDL-treated group. However, the food intake in CRDL-treated groupwas increased to the same amount as the control group in the second weekand maintained at the level similar to the control group through therest of treatment times. FIG. 12B shows the changes of body weightduring the treatment period. Interestingly, while the control groupgained 31% of their body weight from 260.5 g to 341.5 g during the4-weeks fed high fat and high cholesterol diet, the body weights ofWister rats in CRDL-treated group have maintained constant under thesame high fat and high cholesterol diet through the treatment duration.The ability of CRDL to activate AMPK signaling pathway to increaseenergy expenditure and to reduce fat accumulation likely contributes tothis weight reducing effect.

In-vitro Study Results

Example 16 Upregulation of LDLR mRNA Expression in Human HepatomaDerived Cell Line HepG2 by 6 Active Compounds Derived from CorydalisGenus that are all d-(+) Enantiomers

A: Compound Structures

B: Biological Activity

HepG2 cells obtained from American Tissue Culture Collection (Manassas,Va., USA) were seeded in 6-well culture plates at a density of 0.8×10⁶cells/well cultured in EMEM containing 0.5% FBS and were treated witheach purified compound at indicated doses for 8 hours. Total RNA wasisolated, and 2 μg per sample was reverse transcribed with randomprimers using M-MLV (Promega) at 37° C. for 1 hour. PCR was carried outat 94° C. for 30 sec, 60° C. for 30 sec, and 72° C. for 30 sec withinitial activation of the enzyme at 94° C. for 1 minute. Thirty cycleswere performed for LDLR and GAPDH. PCR was performed using primersHLDLR-up and HLDLR-lo for LDLR and primers HGAPDH-up and HGAPDH-lo forGAPDH. The PCR products were separated on a 1% agarose gel and the bandintensity was quantitated. LDLR mRNA levels were corrected by measuringGAPDH mRNA levels.

The potent and dose-dependent effects of (+)-CLMD, 14R-(+)-CRPM,14R,13S-(+)-CRDL, and 14R-(+)-THP on LDLR mRNA expression in HepG2 cellsby a semi-quantitative RT-PCR analysis are shown in FIG. 3.

Specific stereochemical requirements of +/−THP in the upregulation ofLDLR mRNA expression were determined by a similar experiment. HepG2cells were treated with the pure 14R-(+)-THP or the pure 14S-(−)-THP atthe indicated concentrations for 24 hours. The levels of LDLR mRNA andGAPDH mRNA in untreated and the compound-treated HepG2 cells wereassessed by semi-quantitative RT-PCR. The results are as shown in FIG.4.

In yet another experiment, HepG2 cells were treated with variouscompounds for 24 hours at the indicated doses. Total RNA was isolatedand 2 μg was used to generate cDNA in a reaction containing randomprimers and M-MLV at 37° C. for 1 hour in a volume of 25 μl. Real-timePCR was performed on the cDNA using MCEP REALPLEX 2 SYSTEM (Eppendorf)and Universal MasterMix (Applied Biosystems). Human LDLR and GAPDHPre-Developed TaqMan Assay Reagents (Applied Biosystems) were used toassess the levels of mRNA expressions in HepG2. The levels of LDLR mRNAwere normalized to that of GAPDH. Each RNA samples was assayed intriplicate. The abundance of LDLR mRNA in untreated cells was defined as1, and the amounts of LDLR mRNA from compound-treated cells were plottedrelative to that value in FIG. 13. The data shown are mean±s.d. Theresults showed that all 6 compounds with the specific d-(+) enantiomericconfigurations elevated LDLR mRNA levels in a dose-dependent manner.

Example 17 Stimulation of LDLR Ligand Uptake Activity in HepG2 Cells byCorydalis-Derived Compounds and by Some New Compounds of Formula I, II,III, and IV

HepG2 cells (2×10⁵ cells/well) in 24-well culture plates were treatedwith various compounds at indicated concentrations for 20 hours. Thefluorescent 1.1′-dioctadecyl-3,3,3′,3′-tetramethylindocarbocyaninperchlorate (DiI-LDL) (Biomedical Technologies, Stoughton, Mass.) at aconcentration of 2 μg/mL was added to cells at the end of treatment.After 4 hours, the medium was removed, cells were washed with cold PBS,and were examined immediately under a fluorescent microscope (Nikon) at200× amplification. The fluorescent intensity in compound-treated cellswas compared to that in untreated control cells. The compound activitywas graded as follows: +, slightly increased fluorescent intensity overcontrol; ++ modestly increased fluorescent intensity over control; +++,strongly increased fluorescent intensity over control. The results aresummarized below in Table 11. In these experiments berberine chloridewas used for comparison.

TABLE 11 Activity on DiI-LDL A, compound dose ≦ 10 μM Compound No.uptake B, compound dose ≦ 40 μM Berberine + B d-(+)-THP ++ B d-(+)-CRPM++ A d-(+)-CRDL ++ B d-(+)-EGN + A d-(+)-CLMD + B  1 + A 26 ++ A 30 + A34 ++ A 38 ++ A 41 ++ A 43 ++ A 44 ++ A 45 ++ A 46 ++ A 50 +++ A 56 +++A 69 +++ A

Example 18

Activities of some compounds of Formulas I, II, III, and IV on LDLR mRNAexpression were determined. HepG2 cells were treated with new compoundsindividually at a dose less than or equal to 10 μM for 24 hours. TotalRNA was harvested for quantitative real-time RT-PCR analysis using themethod described in Example 16B. The fold activity was derived bydividing the amount of normalized LDLR mRNA in compound-treated cellsover the amount of LDLR mRNA in untreated control cells. In theseexperiments, berberine chloride was included for comparison. Results areshown in Table 12.

TABLE 12 Activity on LDLR mRNA expression Compound No. (Fold of control)BBR 1.30 ± 0.11  1 1.56 ± 0.86 26 1.29 ± 0.10 30 1.47 ± 0.13 34 1.76 ±0.17 37 2.47 ± 0.07 38 1.82 ± 0.10 41 1.60 ± 0.03 43 1.95 ± 0.18 44 2.20± 0.06 45 2.09 ± 0.11 46 1.80 ± 0.16 47 1.63 ± 0.09 48 1.45 ± 0.15 504.74 ± 0.96 51 3.79 ± 0.33 52 3.63 ± 0.17 53 3.96 ± 0.40 56 3.18 ± 0.6357 2.62 ± 0.17 58 3.90 ± 0.01 59 3.73 ± 0.10 60 3.46 ± 0.75 61 1.33 ±0.53 62 0.69 ± 0.02 63 2.94 ± 0.13 64 2.82 ± 0.45 65 2.58 ± 0.46 66 2.54± 0.06 69 2.21 ± 0.09 85 1.42 ± 0.30 86 2.48 ± 0.62 88 1.93 ± 0.24 891.92 ± 0.40 90 1.21 ± 0.07 91 3.77 ± 0.20 92 1.52 ± 0.07 93 1.42 ± 0.1694 1.22 ± 0.18 95 1.43 ± 0.04 96 1.33 ± 0.05 100  1.44 ± 0.59 101  1.97± 0.00

Example 19 Dual Activations of ERK and AMPK Signaling Pathways by theGenus of Corydalis Derived Active Compounds

HepG2 cells cultured in 60-mm dishes were treated with various compoundsat a concentration of 15 μg/ml for 2 hours. Total cell lysate wasisolated and 50 μg of protein was separated on SDS-PAGE and transferredto nitrocellulose membrane. Western blot was performed usinganti-phosphorylated ERK and anti-phosphorylated AMPK antibodies todetect the activation of ERK and AMPK. The membranes were probed againwith anti-ERK and anti-AMPK antibodies to demonstrate equal loadings ofproteins. The results in FIG. 14 show that the levels of phosphorylatedand activated ERK and AMPK were significantly increased incompound-treated cells as compared to untreated control cells.

Western blot analysis was performed to determine the activation of ERKin HepG2 cells by 14R,13S-(+)-CRDL and 14R-(+)-THP. HepG2 cells weretreated with 20 μg/mL 14R,13S-(+)-CRDL and 14R-(+)-THP respectively for0.5, 2, or 8 hours. Cells were then lysed. After protein quantitationusing the BCA™ protein assay reagent (PIERCE), 50 μg protein from eachsample was subjected to SDS-PAGE, followed by Western blotting usinganti-phosphorylated ERK (Cell Signaling) and subsequently reprobed withantibody against β-actin. Results are as shown in FIG. 5.

FIG. 6 shows Western blot analysis of the induction of ACCphosphorylation by 14R-(+)-THP. HepG2 cells were treated with 20 μg/ml14R-(+)-THP for 0.5, 2, or 8 hours. Cells were lysed, and 50 μg proteinfrom each sample was subjected to SDS-PAGE, followed by Western blottingusing anti-phosphorylated ACC (Cell Signaling) and subsequently reprobedwith antibody against β-actin.

Example 20 Reduction of Cellular Triglyceride Content by CorydalisDerived Active Compounds

HepG2 cells seeded in 12-well plate were untreated or treated withvarious active compounds (20 μg/ml) for 24 hours. After treatment, cellswere washed by Tris Buffer (75 μM NaCl, 50 μM Tris-HCl, pH 7.4) twice,and cellular lipids were extracted by 1 ml hexane:isopropanol (3:2)twice. The extractions were evaporated in a chemical hood overnight.Next day, the dried lipids were dissolved in 100 μl isopropanolcontaining 10% Triton-X100. Ten μl per sample was used for TGmeasurements with the commercial available kit obtained from Stanbio. Tonormalize the TG content with cellular protein, after lipid extraction,the cells were lysed in 0.1N NaOH to determine protein concentrationsusing BCA Protein Assay Reagent (Pierce). The results (FIG. 15) showthat the total cellular TG contents were reduced by the compoundtreatments compared to control. It has been demonstrated that AMPKactivation stimulates energy expenditure and reduces TG synthesis. Theactivated AMPK in compound treated cells may account for the reduced TGcontent in cells that were treated with these compounds.

Example 21 Demonstration of Reduction of Intracellular Triglyceride inHepG2 Cells Treated with Some of the NCEs

Berberine chloride was used for comparison. The amount of TG inuntreated control cells was defined as 100% and the amounts of TG incompound-treated cells were divided to that value. Results are shown inTable 13.

TABLE 13 A, compound Activity on reduction of dose ≦ 10 μM Compoundcellular TG accumulation B, compound No. (% of control) dose ≦ 40 μMControl 100 Berberine 70.9 ± 5.4  B 69 60.4 ± 7.34 A 82 53.2 ± 21.3 B 8363.0 ± 0.90 B 22 53.35 ± 12.68 B 91 71.17 ± 2.23  B

Example 22 Examination of Effects of Statins on LDLR mRNA Expression andCellular TG Contents in HepG2 Cells

HepG2 cells were treated with simvastatin at the indicatedconcentrations for 24 hours. After the treatment, a portion of cells washarvested for total RNA, and another portion of cells was used for TGmeasurement. The LDLR mRNA levels were assessed by semi quantitativeRT-PCR, and the cellular TG content was determined as described above.The results in FIG. 16 show that simvastatin dose-dependently increasedLDLR mRNA levels but failed to lower cellular TG content.

Example 23 Examination of the Combined Effects of Statins and d-(+)-THPon LDLR mRNA Expression

To determine whether the stimulatory effects of statins on LDLR geneexpression could be enhanced by corydalis-derived active compounds,HepG2 cells were untreated or treated with a low dose of simvastatin(Simv) or fluvastatin (Fluv) respectively in the absence or the presenceof d-(+) THP, or treated with low concentrations of d-(+) THP alone for24 hours. Quantitative real-time RT-PCR was conducted, and triplicatewas assayed in each RNA sample. Results are shown in FIG. 17. The dataare mean value of triplicates. These data demonstrate that activities ofstatins on upregulation of LDLR expression were not inhibited by d-(+)THP. Instead, addition of d-(+) THP to statin-treated cells furtherimproved the stimulatory effects of statins on LDLR mRNA expression.These preliminary results suggested that corydalis-derived activecompounds of natural origins or synthetic origins have potentials to beused in combinational therapies with HMG CoA reductase inhibitors totreat hyperlipidemic patients.

Example 24 Effect of Compounds of Invention on PCSK9 mRNA Expression

The effects of compounds of Formula I, II, III, and IV on the inhibitionof PCSK9 mRNA expression was examined (Horton J D, Cohen J C, Hobbs H H.“Molecular biology of PCSK9: its role in LDL metabolism” TRENDS inBiochemical Sciences 2007; 32:71-77; Cameron J, Ranheim T, Kulseth M A,Leren T P, Berge K E. “Berberine decreases PCSK9 expression in HepG2cells” Atherosclerosis, 2008 online publication). HepG2 cells weretreated with new compounds individually at 10 uM dose for 24 hours.Total RNA was harvested for quantitative real-time RT-PCR analysis usingmethod described in Example 16B. The fold activity was derived bydividing the amount of normalized PCSK9 mRNA in compound-treated cellsover the amount of PCSK9 mRNA in untreated control cells. In theseexperiments, berberine chloride (10 uM) was included for comparison (seeFIG. 18). These results showed that these new compounds strongly inhibitthe mRNA expression of PCSK9, thereby providing another means toincrease LDLR expression by reducing PCSK9-mediated degradation of LDLRprotein.

Example 25 Comparisons of Simvastatin and New Compound 91 on LDLR andPCSK9 mRNA Expressions

HepG2 cells were treated with simvastatin or compound 91 at theindicated concentrations for 24 hours. Total RNA was harvested forquantitative real-time RT-PCR analysis using method described inExamples 16B. The fold activity was derived by dividing the amount ofnormalized LDLR or PCSK9 mRNA in compound-treated cells over the amountof LDLR or PCSK9 mRNA in untreated control cells. These results as shownin FIG. 19A and FIG. 19B demonstrated that the new compound 91dose-dependently increases LDLR mRNA expression but inhibits the PCSK9mRNA expression, whereas simvastatin increases both LDLR and PCSK9 mRNAexpression.

Example 26 Assessment of Stability of New Compound 69 Under Cell CultureConditions

HepG2 cells were cultured in 6-well cell culture plate in EMEMcontaining 0.5% FBS overnight. Compound 69+ was added to the culturemedium to a final concentration of 20 μM. Medium was collected at 0(control), 4, 8, or 24 hours after the compound addition.

The collected media were analyzed by LC-MS on a Thermo Fisher ScientificSurveyor HPLC system and LCQ ion trap MS with electrospray ionizationsource. A 50×2. 1 mm Hypersil Gold C18 3 um column was used with a flowrate of 200 uL/min. The method conditions were 98% A (0.1% formic acidin water)/2% B (0.1% formic acid in acetonitrile) then a gradient to 95%B in 10 minutes. The injection volume was 10 uL.

LC-MS shows that a single peak with elution time of ˜7 of HPLC and astrong signal at 466.1 (M⁺+H) of MS were detected in all medium samples.Table 14 shows the peak area of 69 in medium collected at different timepoint.

All together, these results suggest the new compound 69 is fairly stableunder cell culture conditions and that the stability of the compound isa contributing factor to its strong effect on the upregulation of LDLRexpression in HepG2 cells.

TABLE 14 Peak Area m/z 466.1 Sample ID Peak Area m/z 466.1 052008-Ctrl2.38e8 052008-4h 9.67e7 052008-8h 9.22e7 052008-24h 7.03e7

The disclosures of each and every patent, patent application andpublication (for example, journals, articles and/or textbooks) citedherein are hereby incorporated herein by reference in their entirety.Definitions in Also, as used herein and in the appended claims, singulararticles such as “a”, “an” and “one” are intended to refer to singularor plural. While the present invention has been described herein inconjunction with a preferred aspect, a person with ordinary skill in theart, after reading the foregoing specification, can effect changes,substitutions of equivalents and other types of alterations to thecompounds of the invention or salts, pharmaceutical compositions,derivatives, prodrugs, metabolites, tautomers or racemic mixturesthereof as set forth herein. Each aspect described above can also haveincluded or incorporated therewith such variations or aspects asdisclosed in regard to any or all of the other aspects. The presentinvention is also not to be limited in terms of the particular aspectsdescribed herein, which are intended as single illustrations ofindividual aspects of the invention. Many modifications and variationsof this invention can be made without departing from its spirit andscope, as will be apparent to those skilled in the art. Functionallyequivalent methods within the scope of the invention, in addition tothose enumerated herein, will be apparent to those skilled in the artfrom the foregoing descriptions. It is to be understood that thisinvention is not limited to particular methods, reagents, compounds,compositions, labeled compounds or biological systems, which can, ofcourse, vary. It is also to be understood that the terminology usedherein is for the purpose of describing particular aspects only, and isnot intended to be limiting. Thus, it is intended that the specificationbe considered as exemplary only with the breadth, scope and spirit ofthe invention indicated only by the appended claims, definitions thereinand any equivalents thereof.

1. A compound of Formula III,

stereoisomers thereof, tautomers thereof, solvates thereof, andpharmaceutically acceptable salts thereof; wherein R₁ and R₂ areindependently —H, —CH₂)₀₋₆COOR′, —C(O)R″, or a substituted orunsubstituted alkyl, alkenyl, cycloalkyl, cycloalkylalkyl, aryl,aralkyl, heteroaryl, heteroarylalkyl, heterocyclyl, or heterocyclylalkylgroup; or R₁ and R₂ together are a methylene group; R₃ and R₈ areindependently —H, —OH, —Cl, —Br, —F, —I, —CN, —NH₂, —C(O)NH₂, —COOH, ora substituted or unsubstituted alkyl, alkenyl, alkoxy or aralkyl group;R₃′ is —H, or R₃ and R₃′ together are an oxo group; R₄ is —H, halogen,—OR′, —OSO₂R″, —OC(O)R″, —OC(O)OR″, —OC(O)NR′R″, —O-alkylene-NR′R′,—O-alkylene-OSO₂R″, —O-alkylene-S(O)₀₋₂R″, —O-alkylene-NR′SO₂R″,—O-alkylene-N(R′)C(O)R′, or a substituted or unsubstituted alkyl group;R₅ and R₆ are independently —H, halogen, —OH, or a substituted orunsubstituted alkoxy group; or R₄ and R₅ together are a methylenedioxygroup, or R₅ and R₆ together are a methylenedioxy group; R₇ is —H,halogen, —OH, or a substituted or unsubstituted alkyl or alkoxy group;each R′ is independently a hydrogen, or a substituted or unsubstitutedalkyl, alkenyl, cycloalkyl, cycloalkylalkyl, aryl, aralkyl, heteroaryl,heteroarylalkyl, heterocyclyl, or heterocyclylalkyl group; each R″ isindependently a substituted or unsubstituted alkyl, alkenyl, cycloalkyl,cycloalkylalkyl, aryl, aralkyl, heteroaryl, heteroarylalkyl,heterocyclyl, or heterocyclylalkyl group; with the proviso that when R₄is —H, —OH or a C₁₋₄ alkoxy group, then R₅ is not —H, —OH or a C₁₋₄alkoxy group; and when R₁ and R₂ are both —CH₃ or when R₁ and R₂together are a methylene group, then R₅ is not OH and R₄ and R₅ togetherare not a methylenedioxy group; and when R₄ is OC(O)R″, then R₅ is notOC(O)R″ or methoxy.
 2. The compound of claim 1, wherein R₁ and R₂ areindependently —H, —CH₂)₀₋₂COOR′, —C(O)(CH₂)₀₋₂R″, or a unsubstitutedC₁₋₆ alkyl group; or R₁ and R₂ together are a methylene group.
 3. Thecompound of claim 1, wherein, R₁ and R₂ together are a methylene group.4. The compound of claim 1, wherein R₃ and R₃′ are each —H, or R₃ andR₃′ together are an oxo group.
 5. The compound of claim 1, wherein R₄ is—H, —OR′, —OSO₂R″, —OC(O)OR″, —OC(O)NR′R″, —O-alkylene-OSO₂R″, or—O-alkylene-NR′R′.
 6. The compound of claim 1, wherein R₄ is —H, —OH, ora substituted or unsubstituted C₁₋₆ alkoxy, C₇₋₁₄ aralkoxy, —OC(O)—(C₁₋₆alkyl), —OC(O)-(aryl), —OC(O)O-(aryl), —OC(O)—NH-(aryl), —O—(C₂₋₆alkylene)-NH—(C₂₋₆ alkyl), —O—(C₂₋₆ alkylene)-NH-(tetrahydropyran),—O—(C₂₋₆ alkylene)-NH-(thiomorpholine dioxide), —O—(C₂₋₆alkylene)-NH-(piperidinyl), —O—(C₂₋₆ alkylene)-NH-(piperazinyl),—O—(C₂₋₆ alkylene)-NH-(morpholinyl), —O—(C₂₋₆ alkylene)-NH-(aralkyl),—O—(C₂₋₆ alkylene)-NH-(cyclopropyl), —OSO₂—(C₃₋₆ cycloalkyl),—OSO₂-(aryl), O—(C₂₋₆ alkylene)-OSO₂-(aryl), —OSO₂-(aralkyl), —O—(C₂₋₆alkylene)-OSO₂-(heteroaryl), —OSO₂—(C₁₋₆ alkyl), —OSO₂-(pyridyl),—OSO₂-(thiazolyl), —O—(C₂₋₆ alkylene)-NHSO₂-(aryl), —O—(C₂₋₆alkylene)-NHSO₂-(heteroaryl), —O—(C₂₋₆ alkylene)-NHC(O)-(aryl), —O—(C₂₋₆alkylene)-NHC(O)-(heteroaryl), —O—(C₀₋₄ alkyl)pyridyl, —O—(C₀₋₄alkyl)pyrimidinyl, —O—(C₀₋₄ alkyl)morpholinyl, —O—(C₀₋₄alkyl)thiomorpholinyl, —O—(C₀₋₄ alkyl)imidazolyl, —O—(C₀₋₄alkyl)thienyl, —O—(C₀₋₄ alkyl)tetrahydropyranyl, —O—(C₀₋₄alkyl)tetrahydrofuranyl, —O—(C₀₋₄ alkyl)pyrrolidinyl, —O—(C₀₋₄alkyl)piperidinyl, or —O—(C₀₋₄ alkyl)piperazinyl group.
 7. The compoundof claim 1, wherein the 14-position in Formula III is the R-(+)stereochemical configuration.
 8. The compound of claim 1, wherein R₅ isOH or unsubstituted alkoxy and R₆ is H.
 9. The compound of claim 1,wherein R₈ is —H, —OH, —COOH, or an unsubstituted alkyl or—CH₂)₁₋₆-phenyl group.
 10. The compound of claim 1, wherein R₁ and R₂are independently —H, —CH₂)₀₋₂COOR′, —C(O)(CH₂)₀₋₂R″, or a unsubstitutedC₁₋₆ alkyl group; or R₁ and R₂ together are a methylene group; R₃ andR₃′ are each —H, or R₃ and R₃′ together are an oxo group; R₄ is —H, —OH,or a substituted or unsubstituted C₁₋₆ alkoxy, C₇₋₁₄ aralkoxy,—OC(O)—(C₁₋₆ alkyl), —OC(O)-(aryl), —OC(O)O-(aryl), —OC(O)—NH-(aryl),—O—(C₂₋₆ alkylene)-NH—(C₂₋₆ alkyl), —O—(C₂₋₆alkylene)-NH-(tetrahydropyran), —O—(C₂₋₆ alkylene)-NH-(thiomorpholinedioxide), —O—(C₂₋₆ alkylene)-NH-(piperidinyl), —O—(C₂₋₆alkylene)-NH-(piperazinyl), —O—(C₂₋₆ alkylene)-NH-(morpholinyl),—O—(C₂₋₆ alkylene)-NH-(aralkyl), —O—(C₂₋₆ alkylene)-NH-(cyclopropyl),—OSO₂—(C₃₋₆ cycloalkyl), —OSO₂-(aryl), —O—(C₂₋₆ alkylene)-OSO₂-(aryl),—OSO₂-(aralkyl), —O—(C₂₋₆ alkylene)-OSO₂-(heteroaryl), —OSO₂—(C₁₋₆alkyl), —OSO₂-(pyridyl), —OSO₂-(thiazolyl), —O—(C₂₋₆alkylene)-NHSO₂-(aryl), —O—(C₂₋₆ alkylene)-NHSO₂-(heteroaryl), —O—(C₂₋₆alkylene)-NHC(O)-(aryl), —O—(C₂₋₆ alkylene)-NHC(O)-(heteroaryl),—O—(C₀₋₄ alkyl)pyridyl, —O—(C₀₋₄ alkyl)pyrimidinyl, —O—(C₀₋₄alkyl)morpholinyl, —O—(C₀₋₄ alkyl)thiomorpholinyl, —O—(C₀₋₄alkyl)imidazolyl, —O—(C₀₋₄ alkyl)thienyl, —O—(C₀₋₄alkyl)tetrahydropyranyl, —O—(C₀₋₄ alkyl)tetrahydrofuranyl, —O—(C₀₋₄alkyl)pyrrolidinyl, —O—(C₀₋₄ alkyl)piperidinyl, or —O—(C₀₋₄alkyl)piperazinyl group; R₅ and R₆ are independently —H, —OH, or anunsubstituted C₁₋₆ alkoxy group; or R₄ and R₅ together are amethylenedioxy group, or R₅ and R₆ together are a methylenedioxy group;and R₈ is —H, —OH, —COOH, or an unsubstituted alkyl or —(CH₂)₁₋₆-phenylgroup.
 11. The compound of claim 1, wherein R₅ is OH or unsubstitutedalkoxy and R₆ is H.
 12. The compound of claim 1, wherein R₈ is —H, —OH,—COOH, or an unsubstituted alkyl or —CH₂)₁₋₆-phenyl group.
 13. Thecompound of claim 1, wherein R₁ and R₂ are independently —H,—CH₂)₀₋₂COOR′, —C(O)(CH₂)₀₋₂R″, or a unsubstituted C₁₋₆ alkyl group; orR₁ and R₂ together are a methylene group; R₃ and R₃′ are each —H, or R₃and R₃′ together are an oxo group; R₄ is a substituted C₁₋₆ alkoxygroup, or a substituted or unsubstituted C₇₋₁₄ aralkoxy, —OC(O)O-(aryl),—OC(O)—NH-(aryl), —O—(C₂₋₆ alkylene)-NH—(C₂₋₆ alkyl), —O—(C₂₋₆alkylene)-NH-(tetrahydropyran), —O—(C₂₋₆ alkylene)-NH-(thiomorpholinedioxide), —O—(C₂₋₆ alkylene)-NH-(piperidinyl), —O—(C₂₋₆alkylene)-NH-(piperazinyl), —O—(C₂₋₆ alkylene)-NH-(morpholinyl),—O—(C₂₋₆ alkylene)-NH-(aralkyl), —O—(C₂₋₆ alkylene)-NH-(cyclopropyl),—OSO₂—(C₃₋₆ cycloalkyl), —OSO₂-(aryl), —O—(C₂₋₆ alkylene)-OSO₂-(aryl),—OSO₂-(aralkyl), —O—(C₂₋₆ alkylene)-OSO₂-(heteroaryl), —OSO₂—(C₁₋₆alkyl), —OSO₂-(pyridyl), —OSO₂-(thiazolyl), —O—(C₂₋₆alkylene)-NHSO₂-(aryl), —O—(C₂₋₆ alkylene)-NHSO₂-(heteroaryl), —O—(C₂₋₆alkylene)-NHC(O)-(aryl), —O—(C₂₋₆ alkylene)-NHC(O)-(heteroaryl),—O—(C₀₋₄ alkyl)pyridyl, —O—(C₀₋₄ alkyl)pyrimidinyl, —O—(C₀₋₄alkyl)morpholinyl, —O—(C₀₋₄ alkyl)thiomorpholinyl, —O—(C₀₋₄alkyl)imidazolyl, —O—(C₀₋₄ alkyl)thienyl, —O—(C₀₋₄alkyl)tetrahydropyranyl, —O—(C₀₋₄ alkyl)tetrahydrofuranyl, —O—(C₀₋₄alkyl)pyrrolidinyl, —O—(C₀₋₄ alkyl)piperidinyl, or —O—(C₀₋₄alkyl)piperazinyl group; R₅ and R₆ are independently —H, —OH, or anunsubstituted C₁₋₆ alkoxy group; or R₄ and R₅ together are amethylenedioxy group, or R₅ and R₆ together are a methylenedioxy group;and R₈ is —H, —OH, —COOH, or an unsubstituted alkyl or —(CH₂)₁₋₆-phenylgroup.
 14. The compound of claim 1, wherein R₁ and R₂ are independently—H, —CH₃, —CH₂COOH, —CH₂C(O)OCH₂CH₃, allyl, or R₁ and R₂ together are amethylene group; R₃ and R₃′ are each —H, or R₃ and R₃′ together are anoxo group; R₄ is —O(CH₂)₂OH, —OCH₂COOH, —OCH₂COOCH₂CH₃, —O(CH₂)₂COOH,—O(CH₂)₂CH₂Br, —O—(CH₂)₂—NH—(CH₂)₂—N(CH₃)₂, —O—(CH₂)₂—NH—(CH₂)₂—OCH₃,—O—(CH₂)₂—NH—(CH₂)₂-SCH₃, —O—(CH₂)₂—NH-morpholinyl,—O—(CH₂)₂—NH—(CH₂)₃—N(CH₃)₂, —O—(CH₂)₂—NH-benzyl,—O—(CH₂)₂—NH—(CH₂)₃-(thiomorpholine dioxide),—O—(CH₂)₂—NH—(CH₂)₃-morpholinyl, —O—(CH₂)₂—NH—(CH₂)₃-tetrahydropyranyl,—O-pyridyl optionally substituted with one or two substituents selectedfrom the group consisting of C₁₋₄ alkyl, —NO₂, and NH₂,—O—(CH₂)₂—S-phenyl, —OSO₂-naphthyl optionally substituted with di(C₁₋₄alkyl), —OSO₂—CF₃, —OSO₂-thiazolyl optionally substituted withacetamido, —O—(CH₂)₀₋₂SO₂-phenyl wherein the phenyl group is optionallysubstituted with one or two substituents selected from the groupconsisting of methyl, methoxy, fluoro, chloro, trifluoromethyl, andnitro, —OSO₂-cyclopentyl, —OSO₂-thienyl, —OSO₂-benzyl,—CH₂)₂-cyclopropyl, —CH₂)₂-morpholinyl, —(CH₂)₂-imidazolyl,—CH₂)₂-pyrrolidinyl, or —CH₂)₂-piperazinyl group, wherein thepiperazinyl group is optionally substituted with methyl, isopropyl, ormethoxyethyl; R₅ and R₆ are independently —H, —OH, or —OCH₃; and R₈ is—H, methyl, ethyl, —COOH, or benzyl.
 15. A composition comprising acompound of claim 1 and a pharmaceutically acceptable carrier.
 16. Acompound of Formula III,

or stereoisomers thereof, tautomers thereof, solvates thereof, andpharmaceutically acceptable salts thereof; wherein R₁ and R₂ areindependently —H, —CH₂)₀₋₆COOR′, —C(O)R″, or a substituted orunsubstituted alkyl, cycloalkyl, cycloalkylalkyl, alkenyl, aryl,aralkyl, heteroaryl, heteroarylalkyl, heterocyclyl, or heterocyclylalkylgroup; or R₁ and R₂ together are a methylene group; R₃ and R₈ areindependently —H, —OH, —Cl, —Br, —F, —I, —CN, —NH₂, —C(O)NH₂, —COOH, ora substituted or unsubstituted alkyl, alkoxy, alkenyl, or aralkyl group;R₃′ is H, or R₃ and R₃′ together are an oxo group; R₄ is halogen,—O-(substituted alkyl), —O-(substituted and unsubstituted aralkyl),—O-(substituted and unsubstituted heterocyclylalkyl), —O-(substitutedand unsubstituted heteroaryl), —OSO₂R″, —OC(O)OR″, —OC(O)NR′R″,—O-alkylene-NR′R′, —O-alkylene-OSO₂R″, —O-alkylene-S(O)₀₋₂R″,—O-alkylene-NR′SO₂R″, —O-alkylene-N(R′)C(O)R′, or a substituted orunsubstituted alkyl group; R₅ and R₆ are independently —H, halogen, —OH,or a substituted or unsubstituted alkoxy group; or R₄ and R₅ togetherare a methylenedioxy group, or R₅ and R₆ together are a methylenedioxygroup; R₇ is H, halogen, OH, or a substituted or unsubstituted alkyl oralkoxy group; each R′ is independently a hydrogen, or a substituted orunsubstituted alkyl, alkenyl, cycloalkyl, cycloalkylalkyl, aryl,aralkyl, heteroaryl, heteroarylalkyl, heterocyclyl, or heterocyclylalkylgroup; and each R″ is independently a substituted or unsubstitutedalkyl, alkenyl, cycloalkyl, cycloalkylalkyl, aryl, aralkyl, heteroaryl,heteroarylalkyl, heterocyclyl, or heterocyclylalkyl group.
 17. Thecompound of claim 16, wherein R₁ and R₂ are independently —H,—CH₂)₀₋₂COOR′, —C(O)(CH₂)₀₋₂R″, or a unsubstituted C₁₋₆ alkyl group; orR₁ and R₂ together are a methylene group.
 18. The compound of claim 16,wherein, R₁ and R₂ together are a methylene group.
 19. The compound ofclaim 16, wherein R₃ and R₃′ are each —H, or R₃ and R₃′ together are anoxo group.
 20. The compound of claim 16, wherein R₄ is —O-(substitutedalkyl), —O-(substituted and unsubstituted aralkyl), —O-(substituted andunsubstituted heterocyclylalkyl), —O-(substituted and unsubstitutedheteroaryl), —OSO₂R″, —OC(O)OR″, —OC(O)NR′R″, —O-alkylene-OSO₂R″, or—O-alkylene-NR′R′.
 21. The compound of claim 16, wherein R₄ is asubstituted C₁₋₆ alkoxy group, or a substituted or unsubstituted C₇₋₁₄aralkoxy, —OC(O)O-(aryl), —OC(O)—NH-(aryl), —O—(C₂₋₆ alkylene)-NH-(C₂₋₆alkyl), —O—(C₂₋₆ alkylene)-NH-(tetrahydropyran), —O—(C₂₋₆alkylene)-NH-(thiomorpholine dioxide), —O—(C₂₋₆alkylene)-NH-(piperidinyl), —O—(C₂₋₆ alkylene)-NH-(piperazinyl),—O—(C₂₋₆ alkylene)-NH-(morpholinyl), —O—(C₂₋₆ alkylene)-NH-(aralkyl),—O—(C₂₋₆ alkylene)-NH-(cyclopropyl), —OSO₂—(C₃₋₆ cycloalkyl),—OSO₂-(aryl), O—(C₂₋₆ alkylene)-OSO₂-(aryl), —OSO₂-(aralkyl), —O—(C₂₋₆alkylene)-OSO₂-(heteroaryl), —OSO₂—(C₁₋₆ alkyl), —OSO₂-(pyridyl),—OSO₂-(thiazolyl), —O—(C₂₋₆ alkylene)-NHSO₂-(aryl), —O—(C₂₋₆alkylene)-NHSO₂-(heteroaryl), —O—(C₂₋₆ alkylene)-NHC(O)-(aryl), —O—(C₂₋₆alkylene)-NHC(O)-(heteroaryl), —O—(C₀₋₄ alkyl)pyridyl, —O—(C₀₋₄alkyl)pyrimidinyl, —O—(C₀₋₄ alkyl)morpholinyl, —O—(C₀₋₄alkyl)thiomorpholinyl, —O—(C₀₋₄ alkyl)imidazolyl, —O—(C₀₋₄alkyl)thienyl, —O—(C₀₋₄ alkyl)tetrahydropyranyl, —O—(C₀₋₄alkyl)tetrahydrofuranyl, —O—(C₀₋₄ alkyl)pyrrolidinyl, —O—(C₀₋₄alkyl)piperidinyl, or —O—(C₀₋₄ alkyl)piperazinyl group.
 22. The compoundof claim 16, wherein the 14-position in Formula III is the R-(+)stereochemical configuration.
 23. A composition comprising a compound ofclaim 16 and a pharmaceutically acceptable carrier.
 24. A compoundselected from group consisting of:10-methoxy-9-(phenylmethoxy)-5,6,7,8,13,13a-hexahydro-2H-1,3-dioxolano[4,5-g]isoquinolino[3,2-a]isoquinoline10-methoxy-9-{[3-(trifluoromethoxy)phenyl]methoxy}-5,6,7,8,13,13a-hexahydro-2H-1,3-dioxolano[4,5-g]isoquinolino[3,2-a]isoquinoline9-[(3-chlorophenyl)methoxy]-10-methoxy-5,6,7,8,13,13a-hexahydro-2H-1,3-dioxolano[4,5-g]isoquinolino[3,2-a]isoquinoline10-methoxy-5,6,7,8,13,13a-hexahydro-2H-1,3-dioxolano[4,5-g]isoquinolino[3,2-a]isoquinolin-9-ylcyclopentanesulfonate,10-methoxy-5,6,7,8,13,13a-hexahydro-2H-1,3-dioxolano[4,5-g]isoquinolino[3,2-a]isoquinolin-9-ylbenzylsulfonate,10-methoxy-5,6,7,8,13,13a-hexahydro-2H-1,3-dioxolano[4,5-g]isoquinolino[3,2-a]isoquinolin-9-yl4-phenylbenzenesulfonate,10-methoxy-5,6,7,8,13,13a-hexahydro-2H-1,3-dioxolano[4,5-g]isoquinolino[3,2-a]isoquinolin-9-yl4-(tert-butyl)benzenesulfonate,(10-methoxy(5,6,7,8,13,13a-hexahydro-2H-1,3-dioxolano[4,5-g]isoquinolino[3,2-a]isoquinolin-9-yloxy))-N-(methylethyl)carboxamide,10-methoxy-5,6,7,8,13,13a-hexahydro-2H-1,3-dioxolano[4,5-g]isoquinolino[3,2-a]isoquinolin-9-yl4-methoxybenzenesulfonate,(10-methoxy(5,6,7,8,13,13a-hexahydro-2H-1,3-dioxolano[4,5-g]isoquinolino[3,2-a]isoquinolin-9-yloxy))-N,N-dimethylcarboxamide,(10-methoxy(5,6,7,8,13,13a-hexahydro-2H-1,3-dioxolano[4,5-g]isoquinolino[3,2-a]isoquinolin-9-yloxy))-N-[4-(trifluoromethoxy)phenyl]carboxamide,N-(3,5-dimethoxyphenyl)(10-methoxy(5,6,7,8,13,13a-hexahydro-2H-1,3-dioxolano[4,5-g]isoquinolino[3,2-a]isoquinolin-9-yloxy))carboxamide,(10-methoxy(5,6,7,8,13,13a-hexahydro-2H-1,3-dioxolano[4,5-g]isoquinolino[3,2-a]isoquinolin-9-yloxy))-N-(3-methylphenyl)carboxamide,(10-methoxy(5,6,7,8,13,13a-hexahydro-2H-1,3-dioxolano[4,5-g]isoquinolino[3,2-a]isoquinolin-9-yloxy))-N-[3-(trifluoromethyl)phenyl]carboxamide,10-methoxy-5,6,7,8,13,13a-hexahydro-2H-1,3-dioxolano[4,5-g]isoquinolino[3,2-a]isoquinolin-9-ylethoxyformate,10-methoxy-5,6,7,8,13,13a-hexahydro-2H-1,3-dioxolano[4,5-g]isoquinolino[3,2-a]isoquinolin-9-yl2-chloro-4-fluorobenzenesulfonate,10-methoxy-5,6,7,8,13,13a-hexahydro-2H-1,3-dioxolano[4,5-g]isoquinolino[3,2-a]isoquinolin-9-yl2-methylbenzenesulfonate,10-methoxy-5,6,7,8,13,13a-hexahydro-2H-1,3-dioxolano[4,5-g]isoquinolino[3,2-a]isoquinolin-9-yl2-fluorobenzenesulfonate,10-methoxy-5,6,7,8,13,13a-hexahydro-2H-1,3-dioxolano[4,5-g]isoquinolino[3,2-a]isoquinolin-9-yl3-fluorobenzenesulfonate,10-methoxy-5,6,7,8,13,13a-hexahydro-2H-1,3-dioxolano[4,5-g]isoquinolino[3,2-a]isoquinolin-9-yl3,4-difluorobenzenesulfonate,10-methoxy-5,6,7,8,13,13a-hexahydro-2H-1,3-dioxolano[4,5-g]isoquinolino[3,2-a]isoquinolin-9-yl(phenylmethoxy)formate,10-methoxy-5,6,7,8,13,13a-hexahydro-2H-1,3-dioxolano[4,5-g]isoquinolino[3,2-a]isoquinolin-9-ylthiophene-2-sulfonate,10-methoxy-5,6,7,8,13,13a-hexahydro-2H-1,3-dioxolano[4,5-g]isoquinolino[3,2-a]isoquinolin-9-yl4-methylbenzenesulfonate,10-methoxy-5,6,7,8,13,13a-hexahydro-2H-1,3-dioxolano[4,5-g]isoquinolino[3,2-a]isoquinolin-9-yl4-fluorobenzenesulfonate,10-methoxy-5,6,7,8,13,13a-hexahydro-2H-1,3-dioxolano[4,5-g]isoquinolino[3,2-a]isoquinolin-9-yl2,4-difluorobenzenesulfonate,10-methoxy-5,6,7,8,13,13a-hexahydro-2H-1,3-dioxolano[4,5-g]isoquinolino[3,2-a]isoquinolin-9-yl3-methyl benzenesulfonate,(10-methoxy(5,6,7,8,13,13a-hexahydro-2H-1,3-dioxolano[4,5-g]isoquinolino[3,2-a]isoquinolin-9-yloxy))-N-benzamide,10-methoxy-5,6,7,8,13,13a-hexahydro-2H-1,3-dioxolano[4,5-g]isoquinolino[3,2-a]isoquinolin-9-yl3-nitrobenzenesulfonate,2,3,10-trimethoxy-9-(phenylmethoxy)-5,6,7,8,13,13a-hexahydroisoquinolino[3,2-a]isoquinoline,10-methoxy-5,6,7,8,13,13a-hexahydro-2H-1,3-dioxolano[4,5-g]isoquinolino[3,2-a]isoquinolin-9-yl4-nitrobenzenesulfonate,10-methoxy-5,6,7,8,13,13a-hexahydro-2H-1,3-dioxolano[4,5-g]isoquinolino[3,2-a]isoquinolin-9-ylquinoline-8-sulfonate,[3,9,10-trimethoxy-5,8,13,13a-tetrahydro-6H-isoquino[3,2-a]isoquinolin-3-yl]-5-(2-oxo-hexahydro-1H-thieno[3,4-d]imidazol-4-yl)pentanoicester,2-[3,9,10-trimethoxy-5,8,13,13a-tetrahydro-6H-isoquino[3,2-a]isoquinolin-3-yl]-aceticacid,9-benzenesulfonyloxy-10-methoxy-5,8,13,13a-tetrahydro-6H-[1,3]dioxolo[4,5-g]isoquino[3,2-α]isoquinoline,9-methanesulfonyloxy-10-methoxy-5,8,13,13a-tetrahydro-6H-[1,3]dioxolo[4,5-g]isoquino[3,2-α]isoquinoline,9-benzenesulfonyloxy-10-methoxy-5,8,13,13a-tetrahydro-6H-[1,3]dioxolo[4,5-g]isoquino[3,2-α]isoquinolinehydrochloride,9-O-3-(1′-bromo-propyl)-10-methoxy-5,8,13,13a-tetrahydro-6H-[1,3]dioxolo[4,5-g]isoquino[3,2-a]isoquinoline,9-O-2-(1′-hydroxy-ethyl)-10-methoxy-5,8,13,13a-tetrahydro-6H-[1,3]dioxolo[4,5-g]isoquino[3,2-a]isoquinoline,2-[10-methoxy-5,8,13,13a-tetrahydro-6H-[1,3]dioxolo[4,5-g]isoquino[3,2-a]isoquinoline-9-yl]-aceticesate,9-(4-chloro-benzenesulfonyloxy)-10-methoxy-5,8,13,13a-tetrahydro-6H-[1,3]dioxolo[4,5-g]isoquino[3,2-α]isoquinoline,9-(4-trifluoromethyl-benzenesulfonyloxy)-10-methoxy-5,8,13,13a-tetrahydro-6H-[1,3]dioxolo[4,5-g]isoquino[3,2-α]isoquinoline,9-(3,4-dimethoxy-benzenesulfonyloxy)-10-methoxy-5,8,13,13a-tetrahydro-6H-[1,3]dioxolo[4,5-g]isoquino[3,2-α]isoquinoline,9-(4-methylsulfonyl-benzenesulfonyloxy)-10-methoxy-5,8,13,13a-tetrahydro-6H-[1,3]dioxolo[4,5-g]isoquino[3,2-α]isoquinoline,9-O-2-(1′-morpholine-ethyl)-10-methoxy-5,8,13,13a-tetrahydro-6H-[1,3]dioxolo[4,5-g]isoquino[3,2-a]isoquinoline,10-methoxy-5,6,7,8,13,13a-hexahydro-2H-1,3-dioxolano[4,5-g]isoquinolino[3,2-a]isoquinolin-9-yl5-(dimethylamino)naphthalenesulfonate,10-methoxy-5,6,7,8,13,13a-hexahydro-2H-1,3-dioxolano[4,5-g]isoquinolino[3,2-a]isoquinolin-9-yl(trifluoromethyl)sulfonate,2,3,10-trimethoxy-5,6,7,8,13,13a-hexahydroisoquinolino[2,1-b]isoquinolin-9-ylbenzenesulfonate,10-methoxy-5,6,7,8,13,13a-hexahydro-2H-1,3-dioxolano[4,5-g]isoquinolino[3,2-a]isoquinolin-9-yl2-(acetylamino)-4-methyl-1,3-thiazole-5-sulfonate,10-methoxy-9-(3-methyl-5-nitro(2-pyridyl)oxy)-5,6,7,8,13,13a-hexahydro-2H-1,3-dioxolano[4,5-g]isoquinolino[3,2-a]isoquinoline,2,10-dimethoxy-3-(phenylmethoxy)-5,6,7,8,13,13a-hexahydroisoquinolino[2,1-b]isoquinolin-9-ylbenzenesulfonate,10-methoxy-9-(5-nitro(2-pyridyl)oxy)-5,6,7,8,13,13a-hexahydro-2H-1,3-dioxolano[4,5-g]isoquinolino[3,2-a]isoquinoline,10-methoxy-5,6,7,8,13,13a-hexahydro-2H-1,3-dioxolano[4,5-g]isoquinolino[3,2-a]isoquinolin-9-ylthiophene-2-sulfonate hydrochloride,10-methoxy-5,6,7,8,13,13a-hexahydro-2H-1,3-dioxolano[4,5-g]isoquinolino[3,2-a]isoquinolin-9-yl3-fluorobenzenesulfonate hydrochloride,10-methoxy-5,6,7,8,13,13a-hexahydro-2H-1,3-dioxolano[4,5-g]isoquinolino[3,2-a]isoquinolin-9-yl3,4-difluorobenzenesulfonatehydrochloride,6-(10-methoxy(5,6,7,8,13,13a-hexahydro-2H-1,3-dioxolano[4,5-g]isoquinolino[3,2-a]isoquinolin-9-yloxy))-5-methyl-3-pyridylamine,10-methoxy-5,6,7,8,13,13a-hexahydro-2H-1,3-dioxolano[4,5-g]isoquinolino[3,2-a]isoquinolin-9-ylpyridine-3-sulfonate,3-Hydroxy-2,10-dimethoxy-5,6,7,8,13,13a-hexahydroisoquinolino[2,1-b]isoquinolin-9-ylbenzenesulfonate,2,10-Dimethoxy-8-prop-2-enyl-3-prop-2-enyloxy-5,6,7,8,13,13a-hexahydroisoquinolino[2,1-b]isoquinolin-9-ylbenzenesulfonate,10-methoxy-5,6,7,8,13,13a-hexahydro-2H-1,3-dioxolano[4,5-g]isoquinolino[3,2-a]isoquinolin-9-yl3-fluorobenzenesulfonate hydrochloride, and9-O-2-(1′-phenylsulfonyl-ethyl)-10-methoxy-5,8,13,13a-tetrahydro-6H-[1,3]dioxolo[4,5-g]isoquino[3,2-a]isoquinoline.25. A composition comprising a compound of claim 24 and apharmaceutically acceptable carrier.